ES2986559T3 - Indole derivatives as histone demethylase inhibitors - Google Patents

Abstract

This application relates to compounds of formula (I), where A, R, R1 and R2 are as defined in the specification, to pharmaceutical compositions containing such compounds and to their use in therapy. (Automatic translation with Google Translate, no legal value)

Description

DESCRIPTION

Indole derivatives as histone demethylase inhibitors

Cross reference to related requests

This application claims priority to, and the benefit of, U.S. Application No. 62/547,433, filed Aug. 18, 2017.

Technical field

The present application relates to indole derivatives, to pharmaceutical compositions containing said compounds and to their use in therapy.

Background

Alterations in the structural and functional states of chromatin, mainly determined by post-translational modifications of histone components, are involved in the pathogenesis of a variety of diseases. The enzymatic processes that govern these post-translational modifications in nucleosomes have become potential targets for so-called epigenetic therapies (Portela, A. et al. Nat. Biotechnol. 2010, 28, 1057-1068).

The discovery of an increasing number of histone lysine demethylases has highlighted the dynamic nature of the regulation of histone methylation, a key chromatin modification that is involved in eukaryotic genome and gene regulation. Histone lysine demethylases represent attractive targets for epigenetic drugs, since their expression and/or activities are often deregulated in cancer (Varier, R. A. et al. Biochim. Biophys. Acta. 2011, 1815, 75-89). A lysine can be mono-, di- and tri-methylated and each of the modifications, even on the same amino acid, can have different biological effects.

Histone lysine demethylases exert their activity through two different types of mechanisms (Anand, R. et al. J. Biol. Chem. 2007, 282, 35425-35429; Metzger, E. et al. Nat. Struct. Mol. Biol. 2007, 14, 252-254). While histone demethylases contain Jumonji domains, which are iron- and 2-oxoglutarate-dependent oxygenases, di- and trimethylated lysines, flavin-dependent histone demethylases (FADs) catalyze the cleavage of mono- and dimethylated lysine residues. Currently, two FAD-dependent demethylases have been identified: LSD1, also known as KDM1A, and LSD2, also known as KDM1B. (Culhane, J. C. et al. Curr. Opin. Chem. Biol. 2007, 11,561-568, Ciccone, D. N. et al. Nature 2009, 461,415-418).

KDM1A is a constituent in several chromatin remodeling complexes and often associates with the co-repressor protein CoREST. KDM1A specifically removes methyl groups from mono- and di-methyl Lys4 of histone H3, which is a well-characterized mark of gene activation. KDM1A represents an interesting target for epigenetic drugs due to its over-expression in solid and hematological tumors (Schulte, J. H. et al. Cancer Res. 2009, 69, 2065-2071; Lim, S. et al. Carcinogenesis 2010, 31, 512-520; Hayami, S. et al. Int. J. Cancer 2011, 128, 574-586; Schildhaus, H. U. et al. Hum. Pathol. 2011, 42, 1667-1675; Bennani-Baiti, I. M. et al. Hum. Pathol. 2012, 43, 1300-1307). Its overexpression correlates with tumor recurrence in prostate cancer (Kahl, P. et al. Cancer Res. 2006, 66, 11341-11347) and KDM1A has a role in various differentiation processes, such as adipogenesis (Musri, M. M. et al. J. Biol. Chem. 2010, 285 , 30034-30041), musculoskeletal differentiation (Choi, J. et al. Biochem. Biophys. Res. Commun. 2010, 401,327-332) and hematopoiesis (Hu, X. et al. Proc. Natl. Acad. Sci. USA 2009, 106, 10141-10146; Li, Y et al. Oncogene. 2012, 31, 5007-5018). KDM1A is also involved in the regulation of cellular energy expenditure (Hino S. et al. Nat Commun. 2012, doi: 10.1038/ncomms1755), in the regulation of thermogenesis and oxidative metabolism in adipose tissue (Duteil et al. Nat Commun. June 10, 2014; 5:4093. doi: 10.1038/ncomms5093.), in the control of viral gene expression checkpoints in productive and latent infections (Roizman, B. J. Virol. 2011,85, 7474-7482), and more specifically in the control of herpes virus infection (Gu, H. J. Virol. 2009, 83, 4376-4385) and HIV transcription (Sakane, N. et al. PLoS Pathog.

2011, 7(8):e1002184). The role of KDM1A in regulating neural stem cell proliferation (Sun, G. et al. Mol. Cell Biol. 2010, 30, 1997-2005) and in controlling neuritis morphogenesis (Zibetti, C. et al. J. Neurosci.

2010, 30, 2521-2532) suggests its possible involvement in neurodegenerative diseases.

Furthermore, KDM1A has been found to be relevant in the control of other important cellular processes such as DNA methylation (Wang, J. et al. Nat. Genet. 2009, 41 (1 ):125-129), cell proliferation (Scoumanne, A. et al. J. Biol. Chem. 2007, 282, 15471-15475; Cho, H. S. et al. Cancer Res. 2011, 71, 655-660), epithelial mesenchymal transition (Lin, T. et al. Oncogene. 2010, 29, 4896-4904) and chromosome segregation (Lv, S. et al. Eur. J. Cell Biol.

2010, 89, 557-563). Furthermore, KDM1A inhibitors are able to reactivate silenced tumor suppressor genes (Huang, Y et al. Proc. Natl. Acad. Sci. U S A. 2007, 104, 8023-8028; Huang, Y et al. Clin. Cancer Res. 2009, 15, 7217-7228), selectively target cancer cells with pluripotent stem cell properties (Wang, J. et al. Cancer Res. 2011, 71, 7238-7249), as well as reactivate the all-trans-retinoic acid differentiation pathway in acute myeloid leukemia (Schenk, T. et al. Nat Med. 2012, 18, 605-611). Furthermore, KDM1A has a clear role in maintaining the oncogenic potential of the MLL-AF9 translocation in leukemia stem cells (Harris et al. Cancer Cell, 21 (2012), 473-487) as well as in human glioblastoma stem-like tumor propagating cells (Suva et al. Cell 2014, 157, 580-594).

The more recently discovered demethylase KDM1B (LSD2) shows a similar specificity to KDM1A for mono- and di-methylated Lys4 of histone H3. KDM1B, unlike KDM1A, does not bind to CoREST and has not been found so far in any of the protein complexes containing KDM1A (Karytinos, A. et al. J. Biol. Chem. 2009, 284, 17775-17782). In contrast, KDM1B forms active complexes with euchromatic histone methyltransferases G9a and NSD3, as well as with cellular factors involved in transcription elongation. KDM1B has been reported to have a role as a regulator of transcription elongation rather than a transcriptional repressor as proposed for KDM1A (Fang, R. et al. Mol. Cell 2010, 39, 222-233).

KDM1A and KDM1B are flavonoid-dependent proteins that share a FAD coenzyme-binding motif, a SWIRM domain, and an amine oxidase domain, all of which are integral to the enzymatic activity of KDM1 family members. Furthermore, both KDM1A and KDM1B show structural similarity to the monoamine oxidases MAO-A and MAO-B. Indeed, tranylcypromine, an MAO inhibitor used as an antidepressant agent, was found to also inhibit KDM1A. The compound acts as an irreversible inhibitor by forming a covalent adduct with the FAD cofactor. (Lee, M. G. et al. Chem. Biol. 2006, 13, 563; Schmidt, D. M. Z. et al. Biochemistry 2007, 46, 4408). Tranylcypromine analogues and their KDM1A inhibitory activity have been described in Bioorg. Med. Chem. Lett. 2008, 18, 3047–3051, in Bioorg. Med. Chem. 2011, 19, 3709–3716, and in J. Am. Chem. Soc.

2011, 132, 6827-6833. Furthermore, arylcyclopropylamine and heteroarylcyclopropylamine derivatives as inhibitors of the KDM1A, MAO-A and/or MAO-B enzyme are described in US2010/324147, WO2012/045883, WO2013/022047 and WO2011/131576.

Reversible inhibitors of KDM1A are not so common and so far there is no clinical data available for them. Examples of reversible inhibitors are aminothiazoles as described in Med. Chem. Commun. 2013, 4, 1513-1522, a N'-(1-phenylethylidene)-benzohydrazide series (J. Med. Chem. 2013, 56, 9496-9508, WO2013025805), or thienopyrrole derivatives (WO2016/034946). Therefore, there is still a need for additional reversible inhibitors that have useful antitumor properties, suitable selectivity and stability of action, and that show increased activity with respect to specific subclasses thereof.

Patent document EP 2993 175 A1 discloses thienopyrroles as histone demethylase inhibitors.

Summary

In one aspect, the application relates to a compound of formula (I)

as defined in claim 1, wherein A is

Hydrogen Res oL1-R5;

R1, R2 are independently hydrogen, OH, C1-C4 alkyl, C3-C7 cycloalkyl, aryl or heteroaryl;

R3 is hydrogen or C1-C4 alkyl;

L1 is a bond, -(CH2)j-Y- or -(CH2)k-,

jes is an integer from 2 to 6;

is an integer from 1 to 6;

Yes oxygen;

R5 is C1-C4 alkyl or aryl, wherein aryl is optionally substituted with one or two substituents chosen from halogen, C1-C6 alkyl or L2-R6;

L2 is -(CH2)m-; or -(CH2)n-W-(CH2)p-;

R<6>is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl;

m, n, p are, independently, zero or an integer from 1 to 6;

Wes oxygen, NH or CH2;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In some embodiments, R3 is methyl or ethyl.

In one embodiment of the compound of formula (I), above:

Res hydrogen oL1-R5;

R1, R2 are independently hydrogen, OH or C1-C4 alkyl;

R3 is methyl or ethyl;

L1 is a bond, -(CH2)2-Y- or -(CH2)k-;

is an integer from 1 to 4;

Yes oxygen;

R5 is C1-C4 alkyl or phenyl substituted with one or two substituents chosen from L2-R6;

L2 is -W-(CH2)p-;

R<6>is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl;

weight zero or 1;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In one embodiment of the compound of formula (I), above:

Res hydrogen oL1-R5;

R1, R2 are independently H or methyl;

R3 is methyl or ethyl;

L1 is -(CH2)j-Y- or -(CH2)k-;

jes 2;

kes 3;

Yes oxygen;

R5 is methyl or phenyl substituted with one or two substituents chosen from L2-R6;

L2 is -(CH2)n-W-(CH2)p-;

nes 0;

weight 0 or 1;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In one embodiment of the compound of formula (I), above:

ResL1-R5;

R1, R2 are hydrogen;

R3 is methyl or ethyl;

L1 is -(CH2)j-Y- or -(CH2)k-;

jes 2;

kes 3;

Yes oxygen;

R5 is phenyl substituted with one or two substituents chosen from L2-R6;

L2 is -(CH2)n-W-(CH2)p-;

nes 0;

weight 0 or 1;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In one embodiment of the compound of formula (I) above:

ResL1-R5;

R1, R2 are hydrogen;

R3 is methyl;

L1 is -(CH2)j-Y-jes 2;

Yes oxygen;

R5 is phenyl substituted with one L2-R6 substituent;

L2 is -(CH2)n-W-(CH2)p-;

nes 0;

weight 0 or 1;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In some embodiments, R6 is piperidinyl or pyrrolidinyl.

In one embodiment, the compound of formula (I), above, is selected from the group consisting of:

1 H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(5-methyl-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(5-ethyl-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(5-bromo-1 H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-ethylthieno[3,2-b]pyrrole-5-yl)-(1 H-indol-3-yl)methanone;

(1-methylindol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[1-(2-methoxyethyl)indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-(2-phenoxyethyl)indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[2-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-[[(3S)-pyrrolidin-3-yl]methoxy]phenoxy]ethyl]indol-3-yl ]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-[[(3R)-pyrrolidin-3-yl]methoxy]phenoxy]ethyl]indol-3-yl ]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(4-piperidyloxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidyloxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(4-piperidylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

[1-[2-[3-(azepan-4-yloxy)phenoxy]ethyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[1-[2-[4-(azepan-4-yloxy)phenoxy]ethyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(4-piperidylmethoxy)phenyl]propyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone;

[1-[3-[4-(azepan-4-yloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[1-[3-[3,4-bis(4-piperidylmethoxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (4-ethylthieno[3,2-b]pyrrole-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone;

(4-ethylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone;

[5-ethyl-1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; and [5-methyl-1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; or a stereoisomer or a pharmaceutically acceptable salt thereof.

In another aspect, the application relates to a compound of formula (I)

as defined in claim 7, wherein

TO

is

ResL1-R5;

R1, R2 are hydrogen;

R4 is methyl or ethyl;

L1 is -(CH2)j-Y- or -(CH2)k-,

jes 2 -,

kes 3;

Yes oxygen;

R5 is phenyl substituted with one or two substituents chosen from L2-R6;

L2 is -(CH2)n-W-(CH2)p-;

R<6>is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl;

nes 0 ;

weight 0 or 1 ;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is:

(6-methylthieno[2,3-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone; either

(6-methylthieno[2,3-b]pirrol-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone.

In another aspect, the application relates to a pharmaceutical composition comprising a compound as defined above together with a pharmaceutically acceptable excipient and/or diluent.

In one embodiment, the pharmaceutical composition further comprises at least one therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors, or a chemotherapeutic agent.

In one embodiment, the pharmaceutical composition is in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable or infusible liquid, solutions, suspensions, emulsions, suppositories, ointments, creams, lotions, gels, pastes or transdermal delivery devices.

In one embodiment, the pharmaceutical composition further comprises at least one therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors or a chemotherapeutic agent, and is in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable or infusible liquid, solutions, suspensions, emulsions, suppositories, ointments, creams, lotions, gels, pastes or transdermal delivery devices.

In another aspect, the application relates to a compound as defined above for use as a medicament.

In another aspect, the application relates to a compound as defined above for use in a method of treatment and/or prevention of cancer, infectious diseases or a disease characterized by aberration of cellular energy metabolism, such as obesity.

In another aspect, the application relates to a compound as defined above for use in a method of treatment and/or prevention of leukemia, non-small cell lung cancer, hepatocellular carcinoma or glioblastomas.

In one embodiment, the application relates to a compound as defined above for use in a method of treating leukemia, non-small cell lung cancer, hepatocellular carcinoma or glioblastomas.

In one embodiment, the application relates to a compound as defined above for use in a method of preventing leukemia, non-small cell lung cancer, hepatocellular carcinoma or glioblastomas.

In another aspect, the application relates to a compound as defined above for use in a method of treatment and/or prevention of acute myeloid leukemia.

In one embodiment, the application relates to a compound as defined above for use in a method of treating acute myeloid leukemia.

In one embodiment, the application relates to a compound as defined above for use in a method of preventing acute myeloid leukemia.

In some embodiments, the methods further comprise administering a therapeutically effective amount of at least one therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors, or a chemotherapeutic agent.

In another aspect, the application relates to a process for obtaining a compound of formula (I) as defined in claim 1, wherein R is hydrogen, the process comprising the preparation of the acyl chloride of formula A2 by reacting the carboxylic acid of formula A1 with thionyl chloride, and the preparation of the indole anion A4 by reacting the indole A3 with methylmagnesium bromide, and the condensation of the acyl chloride of formula A2 with the indole anion A4 to obtain a compound of formula (I), as shown below:

wherein A, R1, R2 and R3 are as defined in claim 1.

In one embodiment, the application relates to a process for obtaining a compound of formula (I) according to claim 1 or claim 7, wherein ResL1-R5, the process comprising the reaction of a compound of formulaB1 with a compound of formulaB2 in the presence of a base, as shown below:

wherein ResL1-R5 and A, R1, R2, R3, R4, L1 and R5 are as defined in claim 1 or claim 7, and LG is a leaving group. In some embodiments, LG is bromine.

Detailed description

The present application relates to substituted indole derivatives that have highly potent inhibitory activities of the KDM1A enzyme and are selective with respect to monoamine oxidases (MAO), useful in the prevention or therapy of diseases and conditions associated with the activity of histone demethylases.

According to the present application, compounds of formula (I) are provided:

claim 1, wherein

R1, R2 are independently hydrogen, OH, C1-C4 alkyl, C3-C7 cycloalkyl, aryl or heteroaryl;

R3 is hydrogen or C1-C4 alkyl;

L1 is a bond, -(CH2)j-Y- or -(CH2)k-,

is an integer from 2 to 6 (for example, 2, 3, 4, 5, or 6);

is an integer from 1 to 6 (for example, 1,2, 3, 4, 5, or 6);

Yes oxygen;

R5 is C1-C4 alkyl or aryl, wherein aryl is optionally substituted with one or two substituents chosen from halogen, C1-C6 alkyl or L2-R6;

L2 is -(CH2)m- or -(CH2)n-W-(CH2)p-;

R<6>is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl;

m, n, p are, independently, zero or an integer from 1 to 6 (for example, 1, 2, 3, 4, 5, or 6);

Wes oxygen, NH or CH2;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In one embodiment, R3 is methyl or ethyl.

In one embodiment:

Res hydrogen oL1-R5;

R1, R are independently hydrogen, OH or C1-C4 alkyl,

R3 is methyl or ethyl

L1 is a bond, -(CH2)2-Y- or -(CH2)k-k is an integer from 1 to 4 (e.g. 1,2, 3 or 4

Yes oxygen

R5 is C1-C4 alkyl or phenyl substituted with one or two substituents chosen from L2-R6;

L2 is -W-(CH2)p;

R6 is heterocyclic, where the heterocyclyl is optionally substituted with C1-C6 alkyl;pes zero or 1;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

In one embodiment, Res hydrogen.

In one embodiment, ResL1-R5.

In one embodiment, R1 and R2 are both hydrogen.

In one embodiment, R1 and R2 are both methyl.

In one embodiment, R1 and R2 are independently hydrogen or C1-C4 alkyl.

In one embodiment, R1 and R2 are independently hydrogen or methyl.

In one embodiment, L1 is -(CH2)j-Y- or -(CH2)k-.

In one embodiment, L1 is -(CH2)j-Y-.

In one embodiment, L1 is -(CH2)k-.

In one embodiment, j is 2, 3, 4, or 5.

In one embodiment, j is 2.

In one embodiment, j is 3.

In one embodiment, j is 4.

In one embodiment, j is 5.

In one embodiment, k is 1, 2, 3, 4, or 5.

In one embodiment, k is 1.

In one embodiment, k is 2.

In one embodiment, k is 3.

In one embodiment, k is 4.

In one embodiment, k is 5.

In one embodiment, R5 is methyl or phenyl substituted with one or two substituents chosen from L2-R6. In one embodiment, R5 is methyl.

In one embodiment, R5 is phenyl substituted with one or two substituents chosen from L2-R6.

In one embodiment, R5 is phenyl substituted with a substituent chosen from L2-R6.

In one embodiment, R5 is phenyl substituted with two substituents chosen from L2-R6.

In one embodiment, L2 is -(CH2)m-.

In one embodiment, L2 is -(CH2)n- W-(CH2)p-.

In one embodiment, Wes oxygen.

In one embodiment, Wes NH.

In one embodiment, W is CH2.

In one embodiment, m is 0, 1, 2, 3, 4, or 5.

In one embodiment, m is 0.

In one embodiment, m is 1.

In one embodiment, m is 2.

In one embodiment, m is 3.

In one embodiment, m is 4.

In one embodiment, m is 5.

In one embodiment, n is 0, 1, 2, 3, 4, or 5.

In one embodiment, n is 0.

In one embodiment, n is 1.

In one embodiment, n is 2.

In one embodiment, n is 3.

In one embodiment, n is 4.

In one embodiment, n is 5.

In one embodiment, p is 0, 1, 2, 3, 4, or 5.

In one embodiment, p is 0.

In one embodiment, p is 1.

In one embodiment, p is 2.

In one embodiment, p is 3.

In one embodiment, p is 4.

In one embodiment, p is 5.

In one embodiment, R6 is pyrrolidyl, pyrrolidinyl, piperidyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, benzodioxolyl, 2,3-dihydro-benzodioxinyl, benzoxazolyl, azetidyl, azepinyl, and diazapinyl, optionally substituted with C1-C6 alkyl. In one embodiment, R6 is piperidinyl or pyrrolidinyl, optionally substituted with C1-C6 alkyl.

In one embodiment, R6 is piperidinyl, optionally substituted with C1-C6 alkyl.

In one embodiment, R6 is pyrrolidinyl, optionally substituted with C1-C6 alkyl.

In one embodiment, R6 is unsubstituted piperidinyl.

In one embodiment, R6 is unsubstituted pyrrolidinyl.

According to the present application, compounds of formula (I) are also provided.

as defined in claim 7, wherein

TO

is

ResL1-R5;

R1, R2 are hydrogen;

R4 is methyl or ethyl;

L1 is -(CH2)j-Y- or -(CH2)k-,

jes 2 -,

kes 3;

Yes oxygen;

R5 is phenyl substituted with one or two substituents chosen from L2-R6;

L2 is -(CH2)n-W-(CH2)p-;

R<6>is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl;

weight 0 or 1 ;

Wes oxygen;

or a stereoisomer or a pharmaceutically acceptable salt thereof.

The compounds of formula (I) according to claim 1 include:

1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(5-methyl-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(5-ethyl-1 H-i indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(5-bromo-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-ethylthieno[3,2-b]pyrrole-5-yl)-(1H-indol-3-yl)methanone;

(1-methylindol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[1-(2-methoxyethyl)indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-(2-phenoxyethyl)indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[2-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrole-5-yl)-[1-[2-[3-[[(3S)-pyrrolidin-3-yl]methoxy]phenoxy]ethyl]indol-3-yl ]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-[[(3R)-pyrrolidin-3-yl]methoxy]phenoxy]ethyl]indol-3-yl ]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(4-piperidyloxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidyloxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(4-piperidylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidylmethoxy)phenoxy]ethyl]indol-3-yl]methanone;

[1-[2-[3-(azepan-4-yloxy)phenoxy]ethyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[1-[2-[4-(azepan-4-yloxy)phenoxy]ethyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(4-piperidylmethoxy)phenyl]propyl]indol-3-yl]methanone;

(4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone;

[1-[3-[4-(azepan-4-yloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[1-[3-[3,4-bis(4-piperidylmethoxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

(4-ethylthieno[3,2-b]pyrrole-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone;

(4-ethylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone;

[5-ethyl-1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

[5-methyl-1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone;

4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidylmethoxy)phenyl]ethyl]indol-3-yl]methanone hydrochloride; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidyloxy)phenyl]ethyl]indol-3-yl]methanone hydrochloride; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-[2-(4-piperidyl)ethoxy] phenyl]ethyl]indol-3-yl]methanone hydrochloride; or stereoisomers or pharmaceutically acceptable salts thereof.

The compounds of formula (I) according to claim 7 include:

(6-methylthieno[2,3-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone;

(6-methylthieno[2,3-b]pyrrol-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone;

or stereoisomers or pharmaceutically acceptable salts thereof.

In another aspect, the application provides the compounds as defined above for use as a medicament.

In another aspect, the application provides the compounds as defined above for use in a method of treatment and/or prevention of cancer, infectious diseases or a disease characterized by aberration of cellular energy metabolism, such as obesity. Preferably, the compounds are for use in a method of treatment and/or prevention of leukemia (e.g., acute myeloid leukemia), non-small cell lung cancer, hepatocellular carcinoma, or glioblastomas. Still preferably, the glioblastomas are giant cell glioblastoma or gliosarcoma.

In some embodiments, the method further comprises administering a therapeutically effective amount of at least one therapeutic agent selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors, or a chemotherapeutic agent.

Another aspect of the application is a pharmaceutical composition comprising a compound as defined above together with a pharmaceutically acceptable excipient and/or diluent.

In some embodiments, the pharmaceutical composition further comprises at least one other therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors, or a chemotherapeutic agent.

In the present application, the term "aryl" includes groups with aromaticity systems, including "conjugated", or multicyclic with one or more aromatic rings and containing no heteroatoms in the ring structure. Examples include phenyl, naphthalenyl, etc. The term "arylene" refers to the corresponding divalent groups, such as phenylene. In one embodiment, "aryl" represents a mono- or bicyclic aromatic ring system of 6 atoms, or 9 or 10 atoms, respectively. Examples of such aryl are phenyl, indenyl, indanyl and naphthyl, and tetrahydronaphthalenyl. Substituted aryl means that the hydrogen atoms on each carbon atom may independently be substituted with a substituent as defined herein above.

In the present application, "heteroaryl" groups are aryl groups, as defined above, except that they have from one to four heteroatoms in the ring structure, and may also be referred to as "arylheterocycles" or "heteroaromatics." As used herein, the term "heteroaryl" is intended to include a stable aromatic heterocyclic ring, such as a stable 5-, 6-, or 7-membered monocyclic or 7-, 8-, 9-, 10-, 11-, or 12-membered bicyclic aromatic heterocyclic ring consisting of carbon atoms and one or more heteroatoms, for example, 1 or 1-2 or 1-3 or 1-4 or 1-5 or 1-6 heteroatoms, or, for example, 1,2, 3, 4, 5, or 6 heteroatoms, independently selected from the group consisting of nitrogen, oxygen, and sulfur. The nitrogen atom may be substituted or unsubstituted (i.e., N or NR where R is H or other substituents, as defined). The nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., N^O and S(O)p, where p = 1 or 2). It should be noted that the total number of S and O atoms in the aromatic heterocycle is not more than 1.

Examples of heteroaryl groups include pyrrole, furan, thiophene, thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole, oxazole, isoxazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like. The term "heteroarylene" refers to the corresponding divalent groups.

Furthermore, the terms "aryl" and "heteroaryl" include multicyclic aryl and heteroaryl groups, e.g., tricyclic, bicyclic, e.g., naphthalene, benzoxazole, benzodioxazole, benzothiazole, benzimidazole, benzothiophene, quinoline, isoquinoline, naphthyridine, indole, benzofuran, purine, benzofuran, deazapurine, indolizine.

The cycloalkyl, heterocycloalkyl, aryl or heteroaryl ring may be substituted at one or more ring positions (e.g., the ring-forming carbon or heteroatom such as N) with such substituents as described above, for example, alkyl, alkenyl, alkynyl, halogen, hydroxy, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonate, phosphinate, amino (including alkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. Aryl and heteroaryl groups may also be fused or bridged with alicyclic or heterocyclic rings, which are not aromatic, to form a multicyclic system (e.g., tetralin, methylenedioxyphenyl, such as benzo[d][1,3]dioxol-5-yl).

"Heterocyclyl" represents a saturated or partially saturated 4- to 12-membered (e.g. 4, 5, 6, 7, 8, 9, 10, 11 or 12-membered) non-aromatic monocyclic, bicyclic, or spirocyclic ring system containing one, two, or three heteroatoms selected from nitrogen, oxygen, and sulfur and three to eleven carbon atoms (e.g. 3, 4, 5, 6, 7, 8, 9, 10 or 11 carbon atoms). Examples of such heterocycles include, but are not limited to: pyrrolidyl, pyrrolidinyl, piperidyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, benzodioxolyl, 2,3-dihydro-benzodioxinyl, benzoxazolyl, azetidyl, azepinyl, and diazapinyl.

The term "C1-C6 alkyl" refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, having one to six carbon atoms. This term encompasses straight or branched hydrocarbon chain radicals, consisting solely of carbon and hydrogen atoms, having 1, 2, 3, 4, 5 or 6 carbon atoms. The "C1-C6 alkyl" group is preferably a straight or branched C1-C4 alkyl group, more preferably a C1-C2 alkyl group. Suitable examples of C1-C6 alkyl include methyl, ethyl, n-propyl, isopropyl, butyl, tert-butyl, pentyl and hexyl. The term "C1-C4 alkyl" refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, having one to four carbon atoms. This term encompasses radicals of linear or branched hydrocarbon chains, consisting only of carbon and hydrogen atoms, which have 1,2, 3 or 4 carbon atoms.

The term "C3-7 cycloalkyl" refers to a saturated monocyclic hydrocarbon ring system having from three to seven carbon atoms. This term encompasses saturated monocyclic hydrocarbon ring systems having 3, 4, 5, 6, or 7 carbon atoms. Suitable examples of C3-6 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term "halogen" refers to fluoro, chloro, bromo or iodo. "Halogens" are preferably fluoro, chloro or bromo, in particular fluoro or chloro.

The term "leaving group" refers to halogen, preferably chloride, bromide or iodide.

The term "reversible inhibitor" refers to an inhibitory molecular entity that interacts with an enzyme by non-covalent interactions and is capable of associating/dissociating from the enzyme.

Pharmaceutically acceptable salts comprise conventional non-toxic salts obtained by salification of a compound of formula (I) with inorganic acids (e.g. hydrochloric, hydrobromic, sulfuric or phosphoric acids), or with organic acids (e.g. acetic, propionic, succinic, benzoic, sulfanilic, 2-acetoxy-benzoic, cinnamic, mandelic, salicylic, glycolic, lactic, oxalic, malic, maleic, malonic, fumaric, tartaric, citric, p-toluenesulfonic, methanesulfonic, ethanesulfonic or naphthalenesulfonic acids). For reviews on suitable pharmaceutical salts, see Berge S. M. et al., J. Pharm. Sci. 1977, 66, 1-19; Gould P L. Int. J. Pharm 1986, 33, 201-217; and Bighley et al. Encyclopedia of Pharmaceutical Technology, Marcel Dekker Inc, New York 1996, Volume 13, pages 453-497. Other salts, which are not pharmaceutically acceptable, for example the trifluoroacetate salt, may be useful in the preparation of compounds of the present application and these form a further aspect of the application. The application includes within its scope all possible stoichiometric and non-stoichiometric forms of the salts of the compounds of formula (I).

Furthermore, the compounds of formula (I) may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents, such as water, EtOH and the like.

Certain compounds of formula (I) may exist in stereoisomeric forms (e.g. they may contain one or more asymmetric carbon atoms). Individual stereoisomers (enantiomers and diastereomers) and mixtures thereof are included within the scope of the present application. The present application also covers individual isomers of the compounds represented by formula (I) as mixtures with isomers thereof in which one or more chiral centers are inverted.

The application also includes all suitable isotopic variations of a compound of the application. Examples of isotopes that may be incorporated into compounds of the application include isotopes such as 2H, 3H, 13C, 14C, 15N, 17O, 18O, 31P, 32P, 35S, 18F and 36Cl, respectively. Certain isotopic variations of the application, for example, those in which a radioactive isotope such as 3H or 14C is incorporated, are useful in studies of tissue distribution of drugs and/or substrates. In addition, substitution with isotopes such as deuterium 2H may provide certain therapeutic advantages resulting from increased metabolic stability. Isotopic variations of the compounds of the application may generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the examples below using appropriate isotopic variations of suitable reagents.

Another embodiment of the application is a process for obtaining a compound of formula (I) as defined in claim 1, wherein R is hydrogen, the process comprising the preparation of the acyl chloride of formula A2 by reacting the carboxylic acid of formula A1 with thionyl chloride, and the preparation of the indole anion A4 by reacting the indole A3 with methylmagnesium bromide, and the condensation of the acyl chloride of formula A2 with the indole anion A4 to obtain a compound of formula (I), as represented in Scheme A below:

wherein A, R1, R2 and R3 are as defined in claim 1.

Carboxylic acids of formula A1 are known compounds or can be prepared as described in PCT application WO2016/034946 from commercially available ethyl 4H-thieno[3,2-b]pyrrole-5-carboxylate (Fluorochem, Cat. No. 067104) or ethyl 6H-thieno[2,3-b]pyrrole-5-carboxylate (Sigma Aldrich, Cat. No. PHo1l284). Indoles of formula A3 are known compounds.

The formation of the acyl chloride of formula A2 can be carried out in a suitable solvent, such as polar aprotic solvents, for example, tetrahydrofuran, 1,4-dioxane, dimethylformamide, dichloromethane, or mixtures thereof, at a temperature ranging from about 0 °C to reflux and for a time ranging from about 30 minutes to 96 hours. The formation of the indole anion A4 can be carried out in a suitable solvent, such as diethyl ether, preferably at a temperature ranging from about 0 °C to room temperature. Preferably, the reaction is carried out under a nitrogen atmosphere. The coupling reaction of an acyl chloride of formula A2 with a compound of formula A4 is carried out in a suitable solvent, such as diethyl ether, at a temperature ranging from about 0 °C to reflux.

Alternatively, a compound of formula (I) may be obtained according to a Friedel-Crafts reaction by reacting an acyl chloride of formula A2 with an indole of formula A3 in the presence of a Lewis acid, for example, AlCb, ZrCR or diethylaluminum chloride, in a suitable solvent, for example, hexane, dichloromethane, or mixtures thereof.

Compounds of formula (I) according to claim 1 or claim 7, wherein RsL1-R5 and L1 and R5 are as defined above, may be obtained by reacting compounds of formula B1 with compounds of formula B2, wherein LG is a leaving group, for example bromine, in a suitable solvent, for example dimethylformamide or dimethylacetamide, and in the presence of a base, for example sodium hydride, at a temperature ranging from about 0 °C to reflux, as depicted in Scheme B below:

where ResL1-R5, and A, R1, R2, R3, R4, L1, and R5 are as defined above, and LG is a leaving group.

Compounds of formula B1 may be prepared as described in Scheme A. Compounds of formula B2 are known compounds or may be prepared by known methods. For example, a compound of formula C5, where LG and R6 are as defined above, Y and L2 are oxygen, may be obtained by the reaction of a diol derivative of formula C1 with an alcohol of formula C2, which is carried out under standard Mitsunobu reaction conditions, for example, by reaction with triphenylphosphine and diethylazodicarboxylate, at a temperature ranging from about 0 °C to 80 °C, in a suitable solvent, such as tetrahydrofuran or toluene or dichloromethane, for a time ranging from about 30 min to 72 h, to give a compound of formula C3. Reaction of a compound of formula C3 with an alcohol of formula C4 under standard Mitsunobu reaction conditions provides intermediate compound C5 as depicted in Scheme C below:

Scheme C.

where p, R6 and LG are as defined above.

The compounds of formula C1, C2 and C4 are known compounds or can be prepared by known methods.

Alternatively, the reaction of a compound of formula D1 with an alcohol of formula D2 under standard Mitsunobu reaction conditions provides the intermediate compound D3 as depicted in Scheme D below:

The compounds of formula D1 and D2 are known compounds or can be prepared by known methods.

In the event that it is necessary to protect a chemical group of a compound of the present application and/or an intermediate thereof, rather than carrying out the reactions described above, said chemical group can be protected and deprotected according to known methods. A comprehensive discussion of the protection/deprotection steps is provided in Greene and Wuts (Greene, T.W.; Wuts, P.G.M. "Protective Groups in Organic Synthesis", John Wiley & Sons Inc., 2006) and in Kocienski (Kocienski, PJ. "Protecting Groups", George Thieme Verlag, 2005).

The salification of the compounds of formula (I), and the preparation of the compounds of formula (I), free of their salts, can be carried out by known conventional methods.

In view of the mechanisms of action described above, the compounds of the present application are useful in the prevention or treatment of tumor-type diseases, including, but not limited to: acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, myelodysplastic syndromes, multiple myeloma, Hodgkin's disease, non-Hodgkin's lymphomas, cutaneous and peripheral T-cell lymphomas, adult T-cell leukemia, large B-cell lymphoma; breast tumors; lung tumors and pleural mesotheliomas, adenocarcinoma, non-small cell lung carcinoma, small cell lung carcinoma; skin tumors including basal cell carcinomas (basaliomas), melanomas, squamous cell carcinoma, Kaposi sarcoma, keratoacanthomas, osteosarcomas, fibrosarcomas, rhabdomyosarcomas, neuroblastomas, glioblastomas, brain tumors, head and neck cancer, testicular and ovarian tumors, cervical carcinoma, endometrial and prostate tumors (e.g., advanced prostate cancer), thyroid carcinomas (e.g., follicular thyroid cancer), colon cancers (e.g., colon adenocarcinoma, colon adenoma), gastric tumors and gastrointestinal adenocarcinomas, hepatocellular carcinomas, pancreatic carcinomas (e.g., exocrine pancreatic carcinoma), renal tumors, teratocarcinomas, and embryonal carcinomas.

The compounds of the application are also useful in preventing or treating infections, including, but not limited to: infections caused by protozoa, fungi, phytotoxic agents, viruses and parasites, for example, HIV or herpes virus infections.

Furthermore, the compounds of the application are also useful in the prevention or treatment of obesity.

The term "therapeutically effective amount" as used herein refers to an amount of formula (I), composition or pharmaceutical composition thereof effective to treat or prevent an identified disease or condition, or to exhibit a detectable therapeutic or inhibitory effect. The effect may be detected by any assay method known in the art. The precise effective amount for a subject will depend on the subject's body weight, size, and health; the nature and extent of the condition; and the therapeutic or combination of therapeutics selected for administration. Therapeutically effective amounts for a given situation may be determined by routine experimentation that is within the skill and judgment of the clinician.

The compounds of formula (I) may also be used in combination with additional agents, in particular antitumor and differentiation agents, either by separate administrations, or by including the two active ingredients in the same pharmaceutical formulation. Non-exhaustive examples of suitable additional agents include:

a) histone deacetylase inhibitors (for example, but not limited to: SAHA, PXD101, JNJ-26481585, SB939, ITF-2357, LBH589, PCI-24781, valproic acid, butyric acid, MS-275, MGCD0103 and FK-228);

b) retinoid receptor modulators, such as 13-c/s-retinoic acid, 9-c/s-retinoic acid, bexarotene, alitretinoin or tretinoin; vitamin D;

c) antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology, such as alkylating agents (e.g. platinum derivatives such as c/s-platin, carboplatin, oxaliplatin, lobaplatin, satraplatin, nedaplatin, heptaplatin; nitrogen mustard such as chlorambucil, melphalan, chlormethine, cyclophosphamide, ifosfamide, trofosfamide, uramustine, bendamustine, estramustine; busulfan, temozolomide or nitrosoureas); antimetabolites (e.g. antifolates such as aminopterin, methotrexate, pemetrexed, raltitrexed); purines such as cladribine, clofarabine, fludarabine, mercaptopurine, pentostatin, thioguanine; pyrimidines such as capecitabine, cytarabine, fluorouracil, floxuridine, gemcitabine; azacitidine, decitabine; cytosine arabinoside or hydroxyurea; antitumor antibiotics (for example, anthracyclines such as aclarubicin, amrubicin, daunomycin, doxorubicin, epirubicin, idarabicin, valrubicin, zorubicin; mitoxantrone; or Streptomyces antibiotics such as actinomycin, bleomycin, mitomycin, or plicamycin); antimitotic agents (e.g., vinca alkaloids such as vincristine, vinblastine, vindesine, or vinorelbine; taxoids such as docetaxel, paclitaxel, or tesetaxel; epothilones such as ixabepilone) and topoisomerase inhibitors (e.g., epipodophyllotoxins such as etoposide and teniposide; amsacrine, camptothecin, irinotecan, rubitecan, and topotecan);

d) cytostatic agents such as antiestrogens (for example, but not limited to: tamoxifen, toremifene, raloxifene, droloxifene and idoxifene), estrogen receptor down-regulators (for example, but not limited to: fulvestrant), antiandrogens (for example, but not limited to: bicalutamide, flutamide, nilutamide, liarozole or cyproterone acetate), LHRH antagonists or LHRH agonists (for example, but not limited to: goserelin, leuprorelin or buserelin), progestogens (for example, but not limited to, megestrol acetate), aromatase inhibitors (for example, but not limited to: anastrozole, letrozole, vorazole and exemestane) and 5-alpha-reductase inhibitors such as finasteride;

e) agents that inhibit cancer cell invasion (e.g. metalloproteinase inhibitors and inhibitors of urokinase plasminogen activator receptor function);

f) inhibitors of growth factor function, for example, growth factor antibodies, growth factor receptor antibodies (for example, but not limited to: the anti-Erbb2 antibody trastuzumab, the anti-Erbbl antibody cetuximab and panitumumab, the anti-IGF1R antibody figitumumab), farnesyl transferase inhibitors, MEK inhibitors, tyrosine kinase inhibitors and serine/threonine kinase inhibitors, for example, enzastaurin, dasatinib, erlotinib, gefitinib, imatinib, lapatinib, nilotinib, sorafenib, sunitinib, everolimus, sirolimus or temsirolimus;

g) antiangiogenic agents, such as those that inhibit the effects of vascular endothelial growth factor, for example, the anti-vascular endothelial cell growth factor antibody bevacizumab, lenalidomide or thalidomide;

h) cell cycle inhibitors including, for example, CDK inhibitors (e.g. but not limited to: flavopiridol, roscovitine) and other cell cycle checkpoint inhibitors; inhibitors of aurora kinase and other kinases involved in the regulation of mitosis and cytokinase;

i) proteasome inhibitors (for example, but not limited to: lactacystin, bortezomib, epoxomicin);

j) HSP90 inhibitors (for example, but not limited to: AT-13387, KOS-953, KOS-1022, CNF-1010, CNF-2024, SNX 5422, STA-9090, NVP-HSP990, NVP-AUY922, PU-H17 and XL-888)

k) selective COX-2 inhibitors (for example, but not limited to, celecoxib) or non-selective NSAIDs (for example, but not limited to: diclofenac, flurbiprofen, ibuprofen, ketoprofen, or naproxen).

In another aspect, a compound of general formula (I) may be used in combination with radiotherapy. In yet another aspect, a compound of general formula (I) may be administered in combination with standard chemotherapy combinations, such as, but not restricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil), CAF (cyclophosphamide, doxorubicin and 5-fluorouracil), AC (doxorubicin and cyclophosphamide), FEC (5-fluorouracil, epirubicin and cyclophosphamide), ACT or ATC (doxorubicin, cyclophosphamide and paclitaxel) or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil and prednisone).

The application also provides pharmaceutical compositions comprising one or more compounds of formula (I) as defined above, and one or more pharmaceutically acceptable excipients and/or diluents. The pharmaceutical compositions may be chosen based on the requirements of the treatment. Such compositions are prepared by mixing and are suitably adapted for oral or parenteral administration, and as such may be administered in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable or infusible liquid, solutions, suspensions, emulsions, suppositories, ointments, creams, lotions, gels, pastes or transdermal delivery devices.

Tablets and capsules for oral administration are usually presented in unit dose form and contain conventional excipients such as binders, fillers (including cellulose, mannitol, lactose), diluents, tableting agents, lubricants (including magnesium stearate), detergents, disintegrants (for example polyvinylpyrrolidone and starch derivatives such as sodium starch glycolate), colouring agents, flavouring agents and wetting agents (for example sodium lauryl sulphate).

Oral solid compositions may be prepared by conventional mixing, filling or tableting methods. The mixing operation may be repeated to distribute the active ingredient in compositions containing large amounts of fillers. Operations of this type are conventional.

Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or they may be presented as a dry product for reconstitution with water or a suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example, sorbitol, syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminium stearate gel, or hydrogenated edible fats; emulsifying agents such as lecithin, sorbitan monooleate or acacia; non-aqueous vehicles (which may include edible oils) such as almond oil, fractionated coconut oil, oily esters such as esters of glycerol, propylene glycol or ethyl alcohol; preservatives such as methyl or propyl p-hydroxybenzoate or sorbic acid, and if desired, conventional flavourings or colourings. Oral formulations also include conventional slow-release formulations such as enteric-coated tablets or granules.

The pharmaceutical preparation for inhalation administration may be supplied from an insufflator or a pressurized nebulizer container.

For parenteral administration, fluid unit dosages may be prepared containing the compound and a sterile vehicle. The compound may be suspended or dissolved, depending on the vehicle and concentration. Parenteral solutions are typically prepared by dissolving the compound in a vehicle, sterilizing by filtration, filling into suitable vials and sealing. Advantageously, adjuvants such as local anesthetics, preservatives and buffering agents may also be dissolved in the vehicle. To enhance stability, the composition may be frozen after the vials have been filled and the water removed under vacuum. Parenteral suspensions are prepared in substantially the same manner, except that the compound may be suspended in the vehicle rather than dissolved, and may be sterilized by exposure to ethylene oxide prior to suspending in the sterile vehicle. Advantageously, a surfactant or wetting agent may be included in the composition to facilitate uniform distribution of the compound of the application.

For buccal or sublingual administration, the compositions may be tablets, lozenges, pastilles or gel.

The compounds may be formulated pharmaceutically as suppositories or retention enemas, for example, containing conventional suppository bases such as cocoa butter, polyethylene glycol, or other glycerides, for rectal administration.

Other means of administration of the compounds of the application relate to topical treatment. Topical formulations may contain, for example, ointments, creams, lotions, gels, solutions, pastes and/or may contain liposomes, micelles and/or microspheres. Examples of ointments include oleaginous ointments such as vegetable oils, animal fats, semi-solid hydrocarbons, emulsifiable ointments, such as hydroxystearin sulfate, anhydrous lanolin, hydrophilic petrolatum, cetyl alcohol, glycerol monostearate, stearic acid, water-soluble ointments containing polyethylene glycols of various molecular weights. Creams, as known to those skilled in the art, are viscous liquids or semi-solid emulsions and contain an oil phase, an emulsifier and an aqueous phase. The oil phase generally contains petrolatum and an alcohol such as cetyl or stearic alcohol. Formulations suitable for topical administration to the eye also include eye drops, wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient.

An additional method of administering the compounds of the application relates to transdermal administration. Typical transdermal formulations comprise conventional aqueous and non-aqueous carriers, such as creams, oils, lotions or pastes, or may be in the form of medicated membranes or patches.

A reference for formulations is Remington's book (''Remington: The Science and Practice of Pharmacy", Lippincott Williams & Wilkins, 2000).

The compounds of the present application may be used alone as sole therapy or in combination with other therapeutic agents for the treatment of the above-mentioned conditions. The combination may be administered as separate (simultaneous, sequential) compositions of the individual components of the treatment or as a single dosage form containing both agents. When the compounds of this application are in combination with other active ingredients, the active ingredients may be formulated separately into single-ingredient preparations in one of the ways described above and then provided as combined preparations, which are given at the same time or at different times, or they may be formulated together into a two- or more-ingredient preparation.

The compounds of general formula (I) may be administered to a patient in a total daily dose of, for example, from 0.001 to 1000 mg/kg of body weight daily. Unit dose compositions may contain such amounts of submultiples thereof to constitute the daily dose. The determination of optimal doses for a particular patient is well known to one skilled in the art.

As is common practice, compositions are usually accompanied by written or printed instructions for use in the treatment in question.

The following Examples are presented to further illustrate the request.

EXAMPLE 1. CHEMICAL SYNTHESIS

Unless otherwise indicated, commercially available reagents and solvents (HPLC grade) were used without further purification. Specifically, the following abbreviations may have been used in descriptions of experimental methods:

Unless otherwise indicated, all temperatures are expressed in °C (degrees Celsius) or K (Kelvin).

H-NMR spectra were acquired with a 500 MHz Varian instrument. Chemical shifts are expressed in parts per million (ppm, units of 5). Coupling constants are expressed in Hertz (Hz) and splitting patterns are described as s (singlet), bs (broad signal), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet).

LC-MS analyses were carried out on a Waters Acquity UPLC or Waters Acquity UPLC H-Class column connected to a SQD single quadrupole (Waters) using an Acquity UPLC BEH C18 (50 x 2.1 mm, 1.7 |jm) or Acquity UPLC HSS T3 (50 x 2.1 mm, 1.8 |jm) column. Phase A consisted of Milli-Q water/CH3CN 95/5 (vol/vol) 0.07% formic acid (by volume) or Milli-Q water 0.07% formic acid (by volume); phase B consisted of CH3CN 0.05% formic acid (by volume); flow rate: 0.6 mL/min; UV detection (DIODE array) from 210 to 400 nm; ESI+ detection in the range of 100-2000 m/z. Yields were calculated assuming products were 100% pure unless otherwise stated.

Intermediate compound 1: 4-methylthieno[3,2-b]pyrrole-5-carboxylic acid

4- Methylthieno[3,2-methylpyrrole-5-carboxylate]ethyl ester

1.5 g (7.7 mmol) of ethyl 4H-thieno[3,2-b]pyrrole-5-carboxylate (Fluorochem, Cat No. 067104) was added at RT in portions to a suspension of 0.46 g (12 mmol) of NaH in 35 mL of dry DMF. After stirring for 20 min at RT, 3.3 g (23 mmol) of CH3I was added in one portion and the mixture was stirred for an additional 30 min. The reaction mixture was then poured into saturated NH4Cl solution and extracted with Et2O. The combined organic layers were dried over Na2SO4, filtered, and the solvent was evaporated to give 1.6 g of ethyl 4-methylthieno[3,2-b]pyrrole-5-carboxylate (99%) as a brown oil. 1H NMR (CDCb) 5 (ppm): 7.34 (d, J=5.4 Hz, 1 H), 7.20 (s, 1 H), 6.95 (d, J=5.4 Hz, 1 H), 4.34 (c, J=7.3 Hz, 2 H), 4.07 (s, 3 H), 1.39 (t, J=7.1 Hz, 3 H); MS (ESI): m/z: 210 [M+H]+

4-Methylthieno[3,2-methylpyrrole-5-carboxylate

0.92 g (38 mmol) of LiOH in 11 mL of H2O at RT was added to a solution of 1.6 g (7.7 mmol) of ethyl 4-methylthieno[3,2-b]pyrrole-5-carboxylate in 11 mL of EtOH. The mixture was stirred for 30 min under reflux. The solvent was evaporated, then H2O was added and the solution was brought to a pH value of about 2 with 2 M HCl. The mixture was extracted with EtOAc, the combined organic phases were dried over Na2SO4 and the solvent was evaporated to give 1.39 g of 4-methylthieno[3,2-b]pyrrole-5-carboxylate (99%) as a beige solid. 1H NMR (CDCb) 5 (ppm): 12.45 (s a, 1 H), 7.54 (d, J=5.4 Hz, 1 H), 7.20 (d, J=5.4 Hz, 1 H), 7.12 (s, 1 H), 3.99 (s, 3 H); MS (ESI): m/z: 182 [M+H]+.

Intermediate compound 2: 4-ethylthieno[3,2-b]pyrrole-5-carboxylic acid

4-Ethylthienoph3,2-bmirrol-5-carboxylate

1.00 g (5.12 mmol) of ethyl 4H-thieno[3,2-b]pyrrole-5-carboxylate was added at RT to a suspension of 0.31 g (7.7 mmol) of NaH in 50 mL of dry DMF. After stirring for 20 min at RT, 2.39 g (15.4 mmol) of ethyl iodide was added and the mixture was stirred for another 30 min at RT. The reaction mixture was then poured into saturated NH4Cl solution and extracted with Et2O. The combined organic layers were dried over Na2SO4 and the solvent was evaporated to give 1.10 g (96%) of ethyl 4-ethylthieno[3,2-b]pyrrole-5-carboxylate as a brown oil. 1H NMR (CDCls) 5 (ppm): 7.34 (d, J=5.4 Hz, 1 H), 7.20 (s, 1 H), 6.96 (d, J=5.4 Hz, 1 H), 4.56 (c, J=7.3 Hz, 2 H), 4.34 (c, J=7.3 Hz, 2 H), 1.46-1.34 (m, 6H); MS (ESI): m/z: 224 [M+H]+.

4-Ethylthieno[3,2-blpyrrole-5-carboxylic acid

0.21 g (9.0 mmol) of LiOH in 4 mL of H2O at RT was added to a solution of 0.40 g (1.8 mmol) of ethyl 4-ethylthieno[3,2-b]pyrrole-5-carboxylate in 4 mL of EtOH. The mixture was stirred for 30 min at reflux, then EtOH was evaporated, water was added and the pH was brought to ca. 2 with 2 M HCl. The mixture was extracted with EtOAc, the combined organic layers were dried over Na2SO4 and the solvent was evaporated to give 0.35 g (quantitative) of 4-ethylthieno[3,2-b]pyrrole-5-carboxylic acid as a beige solid. 1H NMR (DMSO-d6) 5 (ppm): 12.47 (s a, 1 H), 7.56 (d, J=5.4 Hz, 1 H), 7.24 (d, J=5.4 Hz, 1 H), 7.13 (s, 1 H), 4.51 (c, J=7.3 Hz, 2 H), 1.28 (t, J=7.3 Hz, 3H); MS (ESI): m/z: 196 [M+H]+.

Intermediate compound 3: 6-Methylthieno[2,3-b]pyrrole-5-carboxylic acid

6-Methylthienoic acid 2,3-methyl-5-carboxylic acid ethyl ester

Ethyl 6-methylthieno[2,3-b]pyrrole-5-carboxylate was obtained as a yellow solid from ethyl 6H-thieno[2,3-b]pyrrole-5-carboxylate (Eras J. et al. J. Het. Chemistry 1984, 21, 215-217) according to the procedure for Intermediate 1, step 1. 1H NMR (CDCla) 5 (ppm): 7.34 (d, J=5.4 Hz, 1 H), 7.20 (s, 1 H), 6.95 (d, J=5.4 Hz, 1 H), 4.34 (c, J=7.3 Hz, 2 H), 4.07 (s, 3 H), 1.39 (t, J=7.1 Hz, 3 H); MS (ESI): m/z: 210 [M+H]+

6-Methylthienof2.3-blDirrole-5-carboxylic acid

6-Methylthieno[2,3-b]pyrrole-5-carboxylic acid was obtained as a white solid from ethyl 6-methylthieno[2,3-b]pyrrole-5-carboxylate according to the procedure for Intermediate 1, step 2. 1H NMR (DMSO-d6) 5 (ppm): 12.49 (bs, 1 H), 7.17 (d, J = 5.4 Hz, 1 H), 7.08-7.00 (m, 2 H), 3.95 (s, 3 H); MS (ESI): m/z: 182 [M+H]+

Compound 1: 1H-Indole-3-yl-(4-methylthieno[3,2-b]pyrrole-5-yl)methanone

0.26 mL (3.5 mmol) of SOCl2 and 3 drops of DMF were added to a solution of 0.49 g (2.70 mmol) of 4-methylthieno[3,2-b]pyrrole-5-carboxylic acid (Intermediate 1) in 9 mL of THF and the mixture was heated under reflux for about 3 h. The solvent was then evaporated and the acyl chloride formed was used in the next step without further purification.

0.972 mL of a 3 M solution of methylmagnesium bromide in Et2O was slowly added to an ice-cold solution of 0.287 g (2.43 mmol) of indole (Sigma-Aldrich cat. no. I3408) in 3.5 mL of Et2O under a nitrogen atmosphere. After the addition, the reaction mixture was allowed to come to RT, stirred for 2 h, then cooled again to 0 °C and a solution of 4-methylthieno[3,2-b]pyrrole-5-carbonyl chloride in 5 mL of Et2O was added. The resulting mixture was warmed to RT and stirred for 2 h, followed by the slow addition of 6 mL of a saturated NH4Cl solution. The mixture was stirred at room temperature for an additional 1 h and then partitioned between CH2Cl2 and water. The combined organic layers were dried over Na2SO4, filtered, concentrated, and purified by column chromatography (eluent: hexane/acetone, 0% to 15% acetone (by volume)) to give 276 mg (yield: 41%) of 1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone as a mustard-colored solid.

1H NMR (DMSO-d6) 5 (ppm): 11.99 (s a, 1 H), 8.25-8.20 (m, 1 H), 8.13 (s, 1 H), 7.58 ( d, J=5.4 Hz, 1 H), 7.52-7.48 (m, 1 H), 7.27 (d, J=5.4 Hz, 1 H), 7.26-7.17 (m, 2 H), 7.16 (s, 1 H), 4.02 (s, 3 H); MS (ESI): m/z: 281 [M+H]+.

The following compounds (see Table 1) were prepared from 4-methylthieno[3,2-b]pyrrole-5-carboxylic acid (Intermediate 1: Compounds 2-4), 4-ethylthieno[3,2-b]pyrrole-5-carboxylic acid (Intermediate 2: Compound 5), or 6-methylthieno[2,3-b]pyrrole-5-carboxylic acid (Intermediate 3: Compound 6) and the appropriate indoles according to the procedure described for Compound 1.

Table 1.

Intermediate compound 4: 3-[[2-(2-Bromoethoxy)phenoxy]metM]pyrroMdm-1-carboxamide tert-butyl ester

3-[(3-Hydroxyphenoxy)methylpyrrolidine-1-carboxylate tert-butyl ester

0.414 g (1.57 mmol) of tert-butyl 3-(bromomethyl)pyrrolidine-1-carboxylate in 1 mL of DMF was added under nitrogen atmosphere to a solution of 0.219 g (1.99 mmol) of catechol (Sigma-Aldrich, Cat. No. 135011) and 0.649 g (1.99 mmol) of Cs2CO3 in 3 mL of dry DMF. The reaction mixture was stirred overnight at 80 °C. The reaction solution was then diluted with brine and the crude product was extracted with EtOAc. The combined organic layers were dried over Na2SO4, the solvent was removed, and the crude mixture was purified by flash chromatography on silica gel (hexane/acetone, 0% to 10% acetone (by volume)) to give 0.112 g of tert-butyl 3-[(2-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate (yield: 30%). 1H NMR (CDCb) 5 (ppm): 6.98-6.83 (m, 4 H), 5.55 (s, 1 H), 4.07-3.98 (m, 2 H), 3.66-3.58 (m, 1 H), 3.55-3.47 (m, 1 H), 3.45-3.35 (m, 1 H), 3.27-3.18 (m, 1 H), 2.78-2.67 (m, 1 H), 2.17-2.05 (m, 1 H), 1.88-1.74 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

tert-Butyl 3-[(2-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate, 0.16 g (0.74 mmol, 0.15 mL) of DIAD (Sigma-Aldrich, Cat. No. 225541), was added dropwise to a solution comprising 0.145 g (0.494 mmol) of tert-butyl 3-[(2-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate, 0.098 g (0.74 mmol, 0.055 mL) of 2-bromoethanol (Sigma-Aldrich, Cat. No. 65586), and 0.20 g (0.74 mmol) of PPh3 (Sigma-Aldrich, Cat. No. T84409) in 5 mL of Dry THF at 0 °C. The solution was allowed to come to RT and stirred overnight. Then, 0.055 mL of 2-bromoethanol, 0.20 mg of PPh3 and 0.153 mL of DIAD were added to the reaction mixture cooled to 0 °C and the mixture was stirred at RT for another 24 h. The solvent was removed and the crude product was purified by flash chromatography on silica gel (hexane/acetone, 0% to 5% acetone (by volume)) to afford 0.113 g of tert-butyl 3-[[2-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate (yield: 57%) as a colorless oil. 1H NMR (CDCb) 5 (ppm): 7.05-6.84 (m, 4 H), 4.32 (t, J=6.4 Hz, 2 H), 4.04-3.93 (m, 2 H), 3.69-3.59 (m, 3 H), 3.52-3.45 (m, 1 H), 3.42-3.33 (m, 1 H), 3.28-3.20 (m, 1 H), 2.82-2.68 (m, 1 H), 2.15-2.05 (m, 1 H), 1.87-1.76 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 5: tert-butyl 3-[[3-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate

3-[(3-Hydroxyphenoxy)methylpyrrolidine-1-carboxylate tert-butyl ester

0.64 g (3.0 mmol, 0.62 mL) of DIAD (Sigma-Aldrich, Cat. No. 225541) was added dropwise to a solution of 0.33 g (3.0 mmol) of resorcinol (Sigma-Aldrich, Cat. No. 398047), 0.40 g (2.0 mmol) of tert-butyl 3-(hydroxymethyl)pyrrolidine-1-carboxylate (Fluorochem, Cat. No. 048620) and 0.80 g (3.0 mmol) of PPh3 (Sigma-Aldrich, Cat. No. T84409) in 20 mL of dry THF at 0 °C. The reaction mixture was allowed to come to RT and stirred overnight. The solvent was then removed and the crude mixture was purified by flash chromatography on silica gel (hexane/acetone, 0% to 15% acetone (by volume)) to give 0.26 g of tert-butyl 3-[(3-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate (yield: 29%). 1H NMR (CDCb) 5 (ppm): 7.18-7.10 (m, 1 H), 6.53-6.39 (m, 3 H), 3.96 3.84 (m, 2 H), 3.63-3.55 (m, 1 H), 3.53-3.44 (m, 1 H), 3.42-3.32 (m, 1 H), 3.26-3.16 (m, 1 H), 2.74-2.61 (m, 1 H), 2.14 2.02 (m, 1 H), 1.85-1.74 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

3-[[3-(2-Bromoethoxy)phenoxymethylpyrrolidine-1-carboxylate tert-butyl ester

0.17 g (0.79 mmol, 0.16 mL) of DIAD (Sigma-Aldrich, Cat. No. 225541) was added dropwise to a solution comprising 0.154 g (0.525 mmol) of tert-butyl 3-[(3-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate, 0.085 g (0.79 mmol, 0.059 mL) of 2-bromoethanol (Sigma-Aldrich, Cat. No. B65586) and 0.209 g (0.79 mmol) of PPh (Sigma-Aldrich, Cat. No. T84409) in 5 mL of dry THF at 0 °C. The solution was allowed to come to RT and stirred overnight. Then, 0.032 mL of 2-bromoethanol, 0.114 mg of PPh3 and 0.089 mL of DIAD were added to the reaction mixture cooled to 0 °C and the mixture was stirred at RT for another 24 h. The solvent was removed and the crude product was purified by flash chromatography on silica gel (hexane/acetone, 0% to 5% acetone (by volume)) to give 0.128 g of tert-butyl 3-[[3-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate (yield: 61%) as a colorless oil. 1H<r>M<n>(CDCb) 5 (ppm): 7.23-7.15 (m, 1 H), 6.56-6.50 (m, 2 H), 6.496.45 (m, 1 H), 4.29 (t,J=6.4Hz, 2 H), 3.96-3.84 (m, 2 H), 3.65 (t, J=6.1 Hz, 2 H), 3.62-3.56 (m, 1 H), 3.53-3.44 (m, 1 H), 3.41-3.33 (m, 1 H), 3.24-3.17 (m, 1 H), 2.74-2.62 (m, 1 H), 2.13-2.02 (m, 1 H), 1.85-1.73 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 6: (3S)-3-[[3-(2-Bromoethoxy)phenoxy]metM]pyrroMdm-1-carboxamide tert-butyl

(3S)-3-f(3-Hydroxyphenoxy)m-ethylpyrrolidine-1-carboxylate tert-butyl ester

0.13 g of tert-butyl (3S)-3-[(3-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.3 g (2.7 mmol) of resorcinol (Sigma-Aldrich, Cat. No. 398047) and 0.38 g (1.8 mmol) of tert-butyl (3S)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (Fluorochem, Cat. No. 048621) (yield: 25%). 1H NMR (CDCb) 5 (ppm): = 7.17-7.08 (m, 1 H), 6.52-6.37 (m, 3 H), 4.85 (s, 1 H), 3.98-3.82 (m, 2 H), 3.66-3.55 (m, 1 H), 3.54-3.44 (m, 1 H), 3.42-3.32 (m, 1 H), 3.26-3.14 (m, 1 H), 2.75-2.61 (m, 1 H), 2.13-2.00 (m, 1 H), 1.87-1.71 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

(3S)-3-ff3-(2-Bromoethoxy)phenoxymethylpyrrolidine-1-carboxylate tert-butyl ester

0.10 g of tert-butyl (3S)-3-[[3-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 2, from 0.13 g (0.44 mmol) of tert-butyl (3S)-3-[(3-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate and 0.088 g (0.67 mmol, 0.05 mL) of 2-bromoethanol (yield: 56%). 1H NMR (CDCb) 5 (ppm): 7.24-7.15 (m, 1 H), 6.57-6.50 (m, 2 H), 6.49-6.45 (m, 1 H), 4.29 (t, J=6.4 Hz, 2 H), 3.97-3.85 (m, 2 H), 3.64 (t, J=6.4 Hz, 2 H), 3.62-3.57 (m, 1 H), 3.53-3.45 (m, 1 H), 3.41-3.33 (m, 1 H), 3.25 3.17 (m, 1 H), 2.72-2.63 (m, 1 H), 2.12-2.03 (m, 1 H), 1.85-1.74 (m, 1H), 1.48 (s, 9H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 7: (3R)-tert-butyl 3-[[3-(2-Bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate

(3R)-3-f(3-Hydroxyphenoxy)methylpyrrolidine-1-carboxylate tert-butyl ester

0.35 g of tert-butyl (3R)-3-[(3-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.5 g (4.5 mmol) of resorcinol (Sigma-Aldrich, Cat. No. 398047) and 0.636 g (3 mmol) of tert-butyl (3R)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (Fluorochem, Cat. No. 048622) (yield: 40%). 1H NMR (CDCla) 5 (ppm): 7.18-7.09 (m, 1 H), 6.51-6.39 (m, 3 H), 5.06 (s, 1 H), 3.95-3.84 (m, 2 H), 3.64-3.55 (m, 1 H), 3.54-3.44 (m, 1 H), 3.42-3.33 (m, 1 H), 3.24-3.16 (m, 1 H), 2.73-2.61 (m, 1 H), 2.13-2.02 (m, 1 H), 1.87-1.73 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

(3R)-3-ff3-(2-Bromoethoxy)phenoxymethylpyrrolidine-1-carboxylate tert-butyl ester

0.16 g of tert-butyl (3R)-3-[[3-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 2, from 0.18 g (0.60 mmol) of tert-butyl (3R)-3-[(3-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate and 0.12 g (0.90 mmol, 0.067 mL) of 2-bromoethanol (yield: 67%). 1H NMR (CDCb) 5 (ppm): 7.23-7.15 (m, 1 H), 6.55-6.50 (m, 2 H), 6.49-6.46 (m, 1 H), 4.29 (t, J=6.4 Hz, 2 H), 3.97-3.84 (m, 2 H), 3.64 (t, J=6.4 Hz, 2 H), 3.62-3.57 (m, 1 H), 3.53-3.45 (m, 1 H), 3.42-3.33 (m, 1 H), 3.25 3.16 (m, 1 H), 2.72-2.61 (m, 1 H), 2.12-2.01 (m, 1 H), 1.86-1.73 (m, 1H), 1.48 (s, 9H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 8: tert-butyl 3-[[4-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate

3-[(4-Hydroxyphenoxy)methylpyrrolidine-1-carboxylate tert-butyl ester

0.073 g of tert-butyl 3-[(4-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.22 g (2.0 mmol) of hydroquinone (Sigma-Aldrich, Cat. No. H9003) and 0.219 g (1 mmol) of tert-butyl 3-(hydroxymethyl)pyrrolidine-1-carboxylate (Fluorochem, Cat. No. 048620) (yield: 25%). 1H NMR (CDCb) 5 (ppm): 6.82-6.73 (m, 4 H), 4.59 (s, 1 H), 3.94-3.80 (m, 2 H), 3.63-3.55 (m, 1 H), 3.53-3.44 (m, 1 H), 3.42-3.32 (m, 1 H), 3.24-3.14 (m, 1 H), 2.70-2.60 (m, 1 H), 2.12-2.00 (m, 1 H), 1.87-1.73 (m, 1 H), 1.47 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

tert-butyl 3-(4-(2-bromoethoxy)phenoxymethylitrolidine-1-carboxylate

0.053 g of tert-butyl 3-[[4-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 2, from 0.073 g (0.25 mmol) of tert-butyl 3-[(4-hydroxyphenoxy)methyl]pyrrolidine n-1-carboxylate and 0.049 g (0.37 mmol, 0.028 mL) of 2-bromoethanol (yield: 53%). 1H Rm N (CDCb) 5 (ppm): 6.92-6.79 (m, 4 H), 4.25 (t, J=6.4 Hz, 2 H), 3.97-3.81 (m, 2 H), 3.62 (t, J=6.1 Hz, 2 H), 3.60-3.56 (m, 1 H), 3.51 3.44 (m, 1 H), 3.41-3.32 (m, 1 H), 3.23-3.16 (m, 1 H), 2.70-2.61 (m, 1 H), 2.11-2.02 (m, 1 H), 1.85-1.74 (m, 1 H), 1.47 (s, 9 H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 9: tert-butyl 4-[3-(2-Bromoethoxy)phenoxy]piperidm-1-carboxamide

4-(3-Hydroxyphenoxy)piperidine-1-carboxylic acid tert-butyl ester

0.64 g (3.0 mmol, 0.62 mL) of DIAD (Sigma-Aldrich, Cat. No. 225541) was added dropwise to a solution of 0.33 g (3.0 mmol) of resorcinol (Sigma-Aldrich, Cat. No. 398047), 0.42 g (2.0 mmol) of tert-butyl 4-hydroxypiperidine-1-carboxylate (Apollo, Cat. No. OR5404) and 0.80 g (3.0 mmol) of PPIi3 (Sigma-Aldrich, Cat. No. T84409) in 20 mL of dry THF at 0 °C. The reaction mixture was allowed to come to RT and stirred overnight. The solvent was then removed and the crude mixture was purified by flash chromatography on silica gel (hexane/acetone, 0% to 15% acetone, v/v) to give 0.217 g of fertilbutyl 4-(3-hydroxyphenoxy)piperidine-1-carboxylate (yield: 37%) as a white solid. 1H NMR (CDCla) 5 (ppm): 7.16-7.11 (m, 1 H), 6.52-6.48 (m, 1 H), 6.46-6.41 (m, 2 H), 4.96 (s a, 1 H), 4.50-4.38 (m, 1 H), 3.75-3.65 (m, 2 H), 3.39-3.29 (m, 2 H), 1.97-1.86 (m, 2 H), 1.81 1.69 (m, 2 H), 1.48 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

4-[3-(2-Bromoethoxy)phenoxylpiperidine-1-carboxylate tert-butyl ester

0.16 g (0.76 mmol, 0.158 mL) of DIAD (Sigma-Aldrich, Cat. No. 225541) was added dropwise to a solution comprising 0.150 g (0.51 mmol) of tert-butyl 4-[(3-hydroxyphenoxy)piperidine-1-carboxylate, 0.100 g (0.76 mmol, 0.057 mL) of 2-bromoethanol (Sigma-Aldrich, Cat. No. B65586) and 0.203 g (0.76 mmol) of PPh3 (Sigma-Aldrich, Cat. No. T84409) in 5 mL of dry THF at 0 °C. The solution was allowed to come to RT and stirred overnight. Then, 0.028 mL of 2-bromoethanol, 0.101 mg of PPh3 and 0.079 mL of DIAD were added to the reaction mixture cooled to 0 °C, and the mixture was stirred at RT for another 24 h. The solvent was removed and the crude product was purified by flash chromatography on silica gel (hexane/acetone, 0% to 5% acetone (by volume)) to give 0.105 g of tert-butyl 4-[3-(2-bromoethoxy)phenoxy]piperidine-1-carboxylate (yield: 51%) as a colorless oil. 1H NMR (CDCb) 5 (ppm): 7.23-7.15 (m, 1 H), 6.58-6.47 (m, 3 H), 4.50-4.43 (m, 1 H), 4.28 (t, J=6.4 Hz, 2 H), 3.75-3.67 (m, 2 H), 3.64 (t, J=6.4 Hz, 2 H), 3.38-3.28 (m, 2 H), 1.96-1.86 (m, 2 H), 1.81-1.69 (m, 2 H), 1.48 (s, 9 H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 10: tert-butyl 4-[4-(2-bromoethoxy)phenoxy]piperidine-1-carboxylate

4-(4-benzyloxyphenoxy)piperidine-1-carboxylic acid tert-butyl ester

0.8 g of tert-butyl 4-(4-benzyloxyphenoxy)piperidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 1, from 0.60 g (3.0 mmol) of 4-benzyloxyphenol (Sigma-Aldrich, Cat. No. 158348) and 0.93 g (4.5 mmol) of tert-butyl 4-hydroxypiperidine-1-carboxylate (Apollo, Cat. No. OR5404) (yield: 70%).

1H NMR (CDCb) 5 (ppm): 7.46-7.42 (m, 2H), 7.42-7.37 (m, 2H), 7.36-7.31 (m, 1H ), 6.93-6.84 (m, 4 H), 5.03 (s, 2 H), 4.38-4.28 (m, 1 H), 3.76-3.68 (m, 2 H), 3.35-3.24 (m, 2 H), 1.95-1.84 (m, 2 H), 1.78-1.67 ( m, 2 H), 1.48 (s, 9 H); MS (ESI): m/z: 328 [M-56+H]+.

4-(4-Hydroxyphenoxy)piperidine-1-carboxylate tert-butyl ester

0.454 g (1.18 mmol) of tert-butyl 4-(4-benzyloxyphenoxy)piperidine-1-carboxylate dissolved in 47 mL of dry EtOH was hydrogenated in an H-Cube apparatus using a 10% Pd/C (wt) cartridge at 25 °C, atmospheric pressure, and a flow rate of 0.5 mL/min for 5 h. The solution was then concentrated to afford 0.34 g of tert-butyl 4-(4-hydroxyphenoxy)piperidine-1-carboxylate (yield: 98%) as a white solid. 1H NMR (CDCb) 5 (ppm): 6.87-6.71 (m, 4 H), 4.58 (s a, 1 H), 4.36-4.27 (m, 1 H), 3.76-3.67 (m, 2 H), 3.37-3.24 (m, 2 H), 1.95-1.82 (m, 2 H), 1.78-1.66 (m, 2 H), 1.48 (s, 9 H); MS (ESI): m/z: 238 [M-56+H]+.

4-[4-(2-Bromoethoxy)phenoxylpiperidine-1-carboxylate tert-butyl ester

0.168 g (0.419 mmol) of tert-butyl 4-[4-(2-bromoethoxy)phenoxy]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 2, from 0.320 g (1.09 mmol) of tert-butyl 4-(4-hydroxyphenoxy)piperidine-1-carboxylate and 0.214 g (1.63 mmol, 0.122 mL) of 2-bromoethanol (yield: 38%). 1H NMR (CDCla) 5 (ppm): 6.92-6.80 (m, 4 H), 4.37-4.31 (m, 1 H), 4.25 (t, J=6.1 Hz, 2 H), 3.76-3.67 (m, 2 H), 3.63 (t, J=6.4 Hz, 2 H), 3.35-3.26 (m, 2 H), 1.95-1.85 (m, 2 H), 1.77-1.67 (m, 2 H), 1.48 (s, 9 H); MS (ESI): m/z: 344 [M-56+H]+.

Intermediate compound 11: tert-butyl 4-[[3-(2-Bromoethoxy)phenoxy]metM]piperidm-1-carboxmate

tert-butyl 4-[(3-hydroxyphenoxy)methyldimethyldimethylamine-1-carboxylate

0.197 g (0.640 mmol) of tert-butyl 4-[(3-hydroxyphenoxy)methyl]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 1, from 0.333 g (3.00 mmol) of resorcinol (Sigma-Aldrich, Cat. No. 398047) and 0.431 g (2.00 mmol) of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (Sigma-Aldrich, Cat. No. 556017) (yield: 32%). 1H NMR (CDCb) 5 (ppm): 7.18-7.09 (m, 1 H), 6.51-6.37 (m, 3 H), 4.86 (s a, 1 H), 4.23-4.10 (m, 2 H), 3.78 (d, J=6.4 Hz, 2 H), 2.80-2.69 (m, 2 H), 2.02-1.90 (m, 1 H), 1.86-1.76 (m, 2 H), 1.48 (s, 9 H), 1.32-1.19 (m, 2 H); MS (ESI): m/z: 252 [M-56+H]+.

4-[[3-(2-Bromoethoxy)phenoxymethylpiperidine-1-carboxylate tert-butyl ester

0.107 g (0.258 mmol) of tert-butyl 4-[[3-(2-bromoethoxy)phenoxy]methyl]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 2, from 0.152 g (0.490 mmol) of tert-butyl 4-[(3-hydroxyphenoxy)methyl]piperidine-1-carboxylate and 0.097 g (0.74 mmol, 0.055 mL) of 2-bromoethanol (yield: 52%). 1H NMR (CDCb) 5 (ppm): 7.23-7.14 (m, 1 H), 6.57-6.45 (m, 3 H), 4.29 (t, J=6.4 Hz, 2 H), 4.21 4.10 (m, 2 H), 3.79 (d, J=6.4 Hz, 2 H), 3.64 (t, J=6.4 Hz, 2 H), 2.81-2.70 (m, 2 H), 2.02-1.90 (m, 1 H), 1.87-1.78 (m, 2 H), 1.48 (s, 9 H), 1.34-1.21 (m, 2 H); MS (ESI): m/z: 358 [M-56+H]+.

Intermediate compound 12: tert-butyl 4-[[4-(2-bromoethoxy)phenoxy]methyl]piperidine-1-carboxylate

tert-butyl 4-(4-hydroxyphenoxy)methylpiperidine-1-carboxylate

0.28 g of tert-butyl 4-[(4-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 1, from 0.50 g (4.5 mmol) of hydroquinone (Sigma-Aldrich, Cat. No. H9003) and 0.65 g (3.0 mmol) of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (Sigma-Aldrich, Cat. No. 556017) (yield: 31%). 1H NMR (CDCls) 5 (ppm): 6.83-6.70 (m, 4 H), 4.58 (s a, 1 H), 4.22-4.10 (m, 2 H), 3.75 (d, J=6.4 Hz, 2 H), 2.81-2.69 (m, 2 H), 2.00-1.88 (m, 1 H), 1.86-1.77 (m, 2 H), 1.48 (s, 9 H), 1.31-1.23 (m, 2 H); MS (ESI): m/z: 252 [M-56+H]+.

4-[[4-(2-Bromoethoxy)phenoxymethylpiperidine-1-carboxylate tert-butyl ester

0.11 g of tert-butyl 4-[[4-(2-bromoethoxy)phenoxy]methyl]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 2, from 0.28 g (0.90 mmol) of tert-butyl 4-[(4-hydroxyphenoxy)methyl]pyrrolidine-1-carboxylate and 0.18 g (1.35 mmol, 0.101 mL) of 2-bromoethanol (yield: 29%). 1H Rm N (CDCl3) 5 (ppm): 6.89-6.79 (m, 4 H), 4.25 (t, J=6.4 Hz, 2 H), 4.20-4.11 (m, 2 H), 3.76 (d, J=6.4 Hz, 2 H), 3.62 (t, J=6.4 Hz, 2 H), 2.81-2.70 (m, 2 H), 2.00-1.88 (m, 1 H), 1.87-1.77 (m, 2 H), 1.47 (s, 9 H), 1.32-1.23 (m, 2 H); MS (ESI): m/z: 358 [M-56+H]+.

Intermediate compound 13: tert-butyl 4-[3-(2-bromoethoxy)phenoxy]azepane-1-carboxylate

4-(3-Hydroxyphenoxy)azepane-1-carboxylic acid tert-butyl ester

0.43 g of tert-butyl 4-(3-hydroxyphenoxy)azepane-1-carboxylate was prepared according to the procedure for Intermediate 9, step 1, from 0.50 g (4.5 mmol) of resorcinol (Sigma-Aldrich, Cat. No. 398047) and 0.65 g (3.0 mmol) of tert-butyl 4-hydroxyazepane-1-carboxylate (Sigma-Aldrich, Cat. No. CDS009029) (yield: 46%). 1H NMR (CDCla and D2O) 5 (ppm): 7.19-7.05 (m, 1 H), 6.52-6.33 (m, 3 H), 4.48-4.32 (m, 1 H), 3.68-3.15 (m, 4 H), 2.16-1.81 (m, 5 H), 1.72-1.61 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 252 [M-56+H]+.

4-[3-(2-Bromoethoxy)phenoxylazepane-1-carboxylate tert-butyl ester

0.13 g of tert-butyl 3-[[4-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate was prepared according to the procedure for Intermediate 9, step 2, from 0.19 g (0.61 mmol) of tert-butyl 4-(3-hydroxyphenoxy)azepane-1-carboxylate and 0.12 g (0.90 mmol, 0.067 mL) of 2-bromoethanol (yield: 51%). 1H NMR (CDCb) 5 (ppm): 7.23 7.14 (m, 1 H), 6.55-6.48 (m, 2 H), 6.48-6.44 (m, 1 H), 4.48-4.38 (m, 1 H), 4.28 (t, J=6.4 Hz, 2 H), 3.64 (t, J=6.1 Hz, 2 H), 3.60-3.20 (m, 4 H), 2.13-1.86 (m, 5 H), 1.71-1.60 (m, 1 H), 1.49 (s, 9 H); MS (ESI): m/z: 358 [M-56+H]+.

Intermediate compound 14: tert-butyl 4-[4-(2-bromoethoxy)phenoxy]azepane-1-carboxylate

4-(4-Benzyloxyphenoxy)azepane-1-carboxylic acid tert-butyl ester

0.74 g of tert-butyl 4-(4-benzyloxyphenoxy)azepane-1-carboxylate was prepared according to the procedure for Intermediate 9, step 1, from 0.60 g (3.0 mmol) of 4-benzyloxyphenol and 0.97 g (4.5 mmol) of tert-butyl 4-hydroxyazepane-1-carboxylate (Sigma-Aldrich, Cat. No. CDS009029) (yield: 62%). 1H NMR (CDCb) 5 (ppm): 7.46-7.42 (m, 2 H), 7.42-7.37 (m, 2 H), 7.36-7.31 (m, 1 H), 6.92-6.80 (m, 4 H), 5.02 (s, 2 H), 4.37-4.26 (m, 1 H), 3.65-3.23 (m, 4 H), 2.09-1.84 (m, 5 H), 1.70-1.59 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 342 [M-56+H]+.

4-(4-Hydroxyphenoxy)azepane-1-carboxylate tert-butyl ester

0.51 g of tert-butyl 4-(4-hydroxyphenoxy)azepane-1-carboxylate was prepared according to the procedure for Intermediate 10, step 2, from 0.71 g (1.8 mmol) of tert-butyl (4-benzyloxyphenoxy)azepane-1-carboxylate (yield: 92%). 1H NMR (CDCl3) 5 (ppm): 6.82-6.72 (m, 4 H), 4.58 (bs, 1 H), 4.36-4.25 (m, 1 H), 3.65-3.22 (m, 4 H), 2.12-1.83 (m, 5 H), 1.70-1.59 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 252 [M-56+H]+.

tert-butyl 4-(4-(2-)bromoethoxy)phenoxylazeoane-1-carboxylate

0.22 g of tert-butyl 4-[4-(2-bromoethoxy)phenoxy]azepane-1-carboxylate was prepared according to the procedure for Intermediate 9, step 2, from 0.354 g (1.15 mmol) of tert-butyl 4-(4-hydroxyphenoxy)azepane-1-carboxylate and 0.227 g (1.73 mmol, 0.129 mL) of 2-bromoethanol (yield: 47%). 1H<r>M<n>(CDCb) 5 (ppm): 6.92 6.76 (m, 4 H), 4.38-4.29 (m, 1 H), 4.25 (t, J=6.4 Hz, 2 H), 3.67-3.22 (m, 6 H), 2.11-1.84 (m, 5 H), 1.70-1.59 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 358 [M-56+H]+.

Intermediate 15: tert-butyl 3-[[4-(3-bromopropyl)phenoxy]methyl]pyrrolidine-1-carboxylate

0.327 g of tert-butyl 4-[4-(3-bromopropyl)phenoxy]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.215 g (1 mmol) of 4-(3-bromopropyl)phenol (US5204018) and 0.300 g (1.50 mmol) of tert-butyl 4-hydroxypiperidine-1-carboxylate (Apollo, Cat. No. OR5404) (yield: 82%). 1H NMR (CDCb) 5 (ppm): 7.17-7.08 (m, 2 H), 6.89-6.81 (m, 2 H), 4.49-4.38 (m, 1 H), 3.78-3.65 (m, 2 H), 3.40 (t, J=6.6 Hz, 2 H), 3.37-3.30 (m, 2 H), 2.73 (t, J=7.3 Hz, 2 H), 2.18-2.10 (m, 2 H), 1.95-1.87 (m, 2 H), 1.80-1.69 (m, 2 H), 1.47 (s, 9 H); MS (ESI): m/z: 342 [M-56+H]+.

Intermediate compound 18: tert-butyl 4-[4-(3-bromopropyl)phenoxy]azepane-1-carboxylate

0.28 g of tert-butyl 4-[4-(3-bromopropyl)phenoxy]azepane-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.20 g (0.91 mmol) of 4-(3-bromopropyl)phenol (US5204018) and 0.29 g (1.4 mmol) of tert-butyl 4-hydroxyazepane-1-carboxylate (Intermediate 14, step 2) (yield: 75 %). 1H NMR (CDCla) 5 (ppm): 7.14-7.07 (m, 2 H), 6.85-6.77 (m, 2 H), 4.45-4.38 (m, 1 H), 3.65-3.24 (m, 6 H), 2.72 (t, J=7.3 Hz, 2 H), 2.18-2.10 (m, 2 H), 2.10-2.02 (m, 1 H), 1.99-1.85 (m, 4 H), 1.70-1.59 (m, 1 H), 1.48 (s, 9 H); MS (ESI): m/z: 356 [M-56+H]+.

Intermediate 19: tert-butyl 4-[[4-(3-bromopropyl)-2-[(1-tert-butoxycarboxy-4-piperidyl)methoxy]phenoxy]methyl]piperidm-1-carboxylate

tert-butyl 4-[[2-[(1-Tert-butoxycarbonyl-4-DÍDeridyl)methoxyl-4-(3-hydroxyDroDyl)phenoxylmethyllDÍDeridin-1-carboxylate

0.079 g (0.47 mmol) of 4-(3-hydroxypropyl)benzene-1,2-diol (Yang, J. et al. Biorg. Med. Chem Lett.) was suspended.

2014, 24, 2680-2684), 0.03 g (0.2 mmol) of Nal and 0.61 g (1.9 mmol) of Cs2CO3 in 1.5 mL of dry DMF under nitrogen atmosphere. 0.46 g (1.6 mmol) of tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (Sigma-Aldrich, Cat. No. 796719) in 0.8 mL of dry DMF was added and the suspension was stirred at 80 °C for 7 h.

The mixture was cooled to RT and an additional portion of Nal (0.014 g, 0.09 mmol), Cs2CO3 (0.23 g, 0.72 mmol) and tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (0.2 g, 0.72 mmol) were added. The resulting mixture was heated at 80 °C overnight, then cooled to RT and saturated aqueous NH4Cl and EtOAc were added. The organic phase was separated, dried over Na2SO4 and purified by column chromatography on silica gel (eluent: hexane/acetone, 0% to 18% acetone (by volume)) to give 148 mg (yield: 56%) of tert-butyl 4-[[2-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]-4-(3-hydroxypropyl)phenoxy]methyl]piperidine-1-carboxylate as a pale yellow oil. 1H NMR (CDCla) 5 (ppm): 6.85-6.69 (m, 3 H), 4.22-4.08 (m, 4 H), 3.86-3.78 (m, 4 H), 3.69 (t, J=7.3 Hz, 2 H), 2.82-2.71 (m, 4 H), 2.65 (t, J=7.3 Hz, 2 H), 2.06-1.94 (m, 2 H), 1.92-1.78 (m, 6 H), 1.48 (s, 9 H), 1.47 (s, 9 H), 1.33-1.19 (m, 4 H); MS (ESI): m/z: 585 [M+Na]+.

tert-butyl 4-íí4-(3-BromoDroDyl)-2-í(1-tert-butoxycarbonyl-4-DiDeridyl)methoxylphenoxylmethyllDiDeridin-1-carboxylate

A solution of 0.11 g (0.31 mmol) of CBr4 in 0.75 mL of CH2O2 was added dropwise at -18 °C to a solution of 0.144 g (0.256 mmol) of tert-butyl 4-[[2-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]-4-(3-hydroxypropyl)phenoxy]methyl]piperidine-1-carboxylate and 0.081 g (0.31 mmol) of PPh3 in 1.75 mL of CH2O2. The reaction mixture was stirred at RT for 4 h. The solution was then concentrated and the residue was purified by column chromatography (eluent: hexane/acetone, 0% to 7% acetone (by volume)) to give 118 mg (yield: 74%) of tert-butyl 4-[[4-(3-bromopropyl)-2-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]phenoxy]methyl]piperidine-1-carboxylate as a pale yellow oil. 1H NMR (CDCla) 5 (ppm): 6.85-6.67 (m, 3 H), 4.24-4.05 (m, 4 H), 3.87-3.75 (m, 4 H), 3.39 (t, J=6.6 Hz, 2 H), 2.83-2.63 (m, 6 H), 2.19-2.10 (m, 2 H), 2.05-1.93 (m, 2 H), 1.88-1.79 (m, 4 H), 1.48 (s, 9 H), 1.47 (s, 9 H), 1.32-1.18 (m, 4 H); MS (ESI): m/z: 647 [M+Na]+.

Intermediate compound 20: tert-butyl 4-[4-(2-bromoethyl)phenoxy]piperidine-1-carboxylate

0.56 g of tert-butyl 4-[4-(2-bromoethyl)phenoxy]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.40 g (2.0 mmol) of 4-(2-bromoethyl)phenol (Fluorochem, Cat. No. 233801) and 0.61 g (3.0 mmol) of tert-butyl 4-hydroxypiperidine-1-carboxylate (Apollo, Cat. No. OR5404) (yield: 73%).

1H NMR (CDCls) 5 (ppm): 7.18-7.08 (m, 2H), 6.91-6.81 (m, 2H), 4.50-4.37 (m, 1H ), 3.76-3.65 (m, 2 H), 3.54 (t, J=7.6 Hz, 2 H), 3.39-3.28 (m, 2 H), 3.11 (t, J=7.6 Hz, 2 H), 1.99-1.85 (m, 2 H), 1.80-1.68 (m, 2 H), 1 .48 (s, 9H); MS (ESI): m/z: 328 [M-56+H]+.

Intermediate compound 21: tert-butyl 4-[[4-(2-BromoethN)phenoxy]metN]piperidm-1-carboxmate

0.39 g of tert-butyl 4-[4-(2-bromoethyl)phenoxy]methyl]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.30 g (1.5 mmol) of 4-(2-bromoethyl)phenol (Fluorochem, Cat. No. 233801) and 0.48 g (2.25 mmol) of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (Sigma-Aldrich, Cat. No. 556017) (yield: 65%). 1H NMR (CDCla) 5 (ppm): 7.17 - 7.09 (m, 2 H), 6.88 - 6.78 (m, 2 H), 4.23 - 4.07 (m, 2 H), 3.79 (d, J = 6.4 Hz, 2 H), 3.54 (t, J = 7.6 Hz, 2 H), 3.11 (t, J = 7.6 Hz, 2 H), 2.80 - 2.70 (m, 2 H), 2.01 - 1.89 (m, 1 H), 1.87 - 1.79 (m, 2 H), 1.47 (s, 9 H), 1.32 - 1.22 (m, 2 H); MS (ESI): m/z: 342 [M-56+H]+.

Intermediate compound 22: tert-butyl 4-[[4-(2-bromoethyl)phenoxy]ethyl]piperidine-1-carboxylate

0.52 g of tert-butyl 4-[4-(2-bromoethyl)phenoxy]ethyl]piperidine-1-carboxylate was prepared according to the procedure for Intermediate 5, step 1, from 0.27 g (1.32 mmol) of 4-(2-bromoethyl)phenol (Fluorochem, Cat. No. 233801) and 0.47 g (1.98 mmol) of tert-butyl 4-(hydroxyethyl)piperidine-1-carboxylate (Sigma-Aldrich, Cat. No. 547247) (yield: 96%). 1H NMR (CDCls) 5 (ppm): 7.17 - 7.09 (m, 2 H), 6.88 - 6.81 (m, 2 H), 4.16 - 4.05 (m, 2 H), 4.00 (t, J = 6.1 Hz, 2 H), 3.54 (t, J = 7.8 Hz, 2 H), 3.11 (t, J = 7.8 Hz, 2 H), 2.76 - 2.65 (m, 2 H), 1.77 - 1.65 (m, 5 H), 1.47 (s, 9 H), 1.23 - 1.12 (m, 2 H); MS (ESI): m/z: 356 [M-56+H]+.

Intermediate compound 23: Tert-butyl-(1H-indol-7-yloxy)-dimethyl-silane

0.307 mL (2.2 mmol) of TEA and 0.01 g (0.8 mmol) of DMAP were added to an ice-cold solution of 0.28 g (2.0 mmol) of 1 H-indol-7-ol in 30 mL of dry CH2Cl2. After 10 min, 0.34 mg (2.2 mmol) of TBDMSCI was added and the mixture was allowed to warm to RT and stirred overnight. Water (50 mL) was then added and the reaction mixture was extracted with CH2O2 (3*100 mL). The combined organic layers were dried over Na2SO4, filtered, concentrated, and purified by column chromatography (eluent: hexane/acetone, 0% to 5% acetone (by volume)) to give 403 mg (yield: 81%) of tert-butyl-(1H-indol-7-yloxy)-dimethyl-silane as a pale yellow oil. 1H NMR (DMSO-da) 5 (ppm): 10.78-10.62 (b s, 1 H), 7.28-7.22 (m, 1 H), 7.15-7.09 (m, 1 H), 6.86-6.79 (m, 1 H), 6.56-6.50 (m, 1 H), 6.41-6.36 (m, 1 H), 1.00 (s, 9 H), 0.25 (s, 6 H); MS (ESI): m/z: 248 [M+H]+.

Intermediate compound 24: [7-[ferc-But¡l(d¡met¡l)s¡l¡l]oxMH-mdol-3-¡l]-(4-met¡lt¡ene[3,2-b ]pyrrole-5-l)methanone

0.13 mL (1.79 mmol) of SOCb and 3 drops of DMF were added to a solution of 0.25 g (1.4 mmol) of 4-methylthieno[3,2-b]pyrrole-5-carboxylic acid (Intermediate 1) in 4 mL of THF and the mixture was heated under reflux for about 3 h. The solvent was then evaporated and the acyl chloride formed was used in the next step without further purification.

0.660 mL of a 300 M solution of methylmagnesium bromide in Et2O was slowly added to an ice-cold solution of 0.376 g (1.52 mmol) of tert-butyl-[1H-indol-7-yloxy]-dimethyl-silane in 3 mL of Et2O under a nitrogen atmosphere. The reaction mixture was allowed to come to RT, stirred for 2 h and then cooled again to 0 °C. A solution of (quantity of) 4-methylthieno[3,2-b]pyrrole-5-carbonyl chloride in 5 mL of Et2O was added and the resulting mixture was warmed to RT and stirred for 2 h followed by slow addition of 6 mL of a saturated NH4Cl solution. The mixture was stirred at RT for an additional 1 h and then partitioned between CH2Cl2 and water. The combined organic layers were dried over Na2SO4, filtered, concentrated, and purified by column chromatography (eluent: hexane/acetone, 0% to 12% acetone (by volume)) to give 160 mg (yield: 28%) of [7-[fert-butyl(dimethyl)silii]oxy-1H-indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone as a beige solid. 1H NMR (CDCb) 5 (ppm): 8.54 (b s, 1 H), 7.99-7.96 (m, 1 H), 7.86-7.83 (m, 1 H), 7.37 (d, J=5.4 Hz, 1 H), 7.18-7.13 (m, 1 H), 7.05 (s, 1 H), 7.02 (d, J=5.4 Hz, 1 H), 6.78-6.73 (m, 1 H), 4.13 (s, 3 H), 1.06 (s, 9 H), 0.32 (s, 6 H); MS (ESI): m/z: 411 [M+H]+.

Intermediate Compound 25: 3-[[2-[2-[3-(4-Methyltene[3,2-b]pyrrole-5-carboml)mdoM-l]ethoxy ]fenox¡]met¡l]pyrrolídin-1-carbox¡late of ferc-butNo

0.034 g (0.12 mmol) of 1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone (Compound 1) was added to a suspension of 0.0058 g (0.15 mmol) of NaH (60% dispersion in mineral oil (by weight)) in dry DMF (0.5 mL) cooled to 0 °C. The mixture was stirred at 0 °C for 20 min, then a solution of 0.080 g (0.15 mmol) of tert-butyl 3-[[2-(2-bromoethoxy)phenoxy]methyl]pyrrolidine-1-carboxylate (Intermediate 4) in 1 mL of dry DMF was added at 0 °C and the mixture was stirred for 5 h at RT. The reaction was quenched by adding saturated aqueous NaHCO3 solution and the product was extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and evaporated to dryness. The crude product was purified by flash chromatography on silica gel (CH2Cl2/acetone, 0% to 4% acetone (by volume)) to afford 0.053 g of tere-butyl 3-[[2-[2-[3-(4-methylthieno[3,2-b]pyrrole-5-carbonyl)indol-1-yl]ethoxy]phenoxy]methyl]pyrrolidine-1-carboxylate (Intermediate 25, yield: 73%) as a pale yellow solid. 1H NMR (CDCb) 5 (ppm): 8.47-8.37 (m, 1 H), 8.02 (s, 1 H), 7.51 7.43 (m, 1 H), 7.39-7.30 (m, 3 H), 7.01 (d, J=5.4 Hz, 1 H), 6.99-6.91 (m, 2 H), 6.90-6.85 (m, 2 H), 6.82-6.77 (m, 1 H), 4.62 (t, J=4.9 Hz, 2 H), 4.39 (t, J=4.9 Hz, 2 H), 4.12 (s, 3 H), 3.82-3.75 (m, 2 H), 3.47-3.40 (m, 1 H), 3.39-3.29 (m, 1 H), 3.26-3.15 (m, 1 H), 3.12-3.02 (m, 1 H), 2.45-2.35 (m, 1 H), 1.88-1.77 (m, 1 H), 1.55-1.51 (m, 1 H), 1.45 (s, 9 H); MS (ESI): m/z: 600 [M+H]+.

The following BOC intermediates were obtained from Intermediates 1, 5 or 6 and the corresponding bromine derivatives according to Intermediate 22 described in the procedure using DMF or DMA as solvent:

Compound 7: (1-MetNmdol-3-N)-(4-metMthieno[3,2-b]pyrrole-5-N)methanone

0.016 g (0.057 mmol) of 1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone (Compound 1) was added to a solution of 0.0027 g (0.068 mmol) of NaH in 1 mL of DMF and the mixture was stirred for 20 min. 0.0043 mL (0.068 mmol) of methyl iodide was added and the resulting mixture was stirred at RT for 30 min and then partitioned between EtOAc and water. The combined organic layers were dried over Na2SO4, filtered, concentrated and purified by column chromatography (eluent: hexane/acetone (hexane/acetone, 85:15, v/v) to give 9 mg (yield: 54%) of (1-methylindol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone as a pale yellow solid. 1H NMR (CDCls) 5 (ppm): 8.48-8.36 (m, 1 H), 7.77 (s, 1 H), 7.44-7.31 (m, 4 H), 7.05-6.98 (m, 2 H), 4.12 (s, 3 H), 3.90 (s, 3 H); MS (ESI): m/z: 295 [M+H]+.

Compound 8: [1-(2-Methoxyethyl)indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone

0.029 g of [1-(2-Methoxyethyl)indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone was prepared from 0.034 g (0.12 mmol) of 1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone (Compound 1) and 0.058 g (0.17 mmol, 0.016 mL) of 1-bromo-2-methoxyethane (Sigma-Aldrich, Cat. No. 238155) according to the procedure described for Compound 7 (yield: 71%). 1H NMR (CDCla) 5 (ppm): 8.49-8.37 (m, 1 H), 7.87 (s, 1 H), 7.44-7.39 (m, 1 H), 7.38-7.30 (m, 3 H), 7.05-6.98 (m, 2 H), 4.37 (t, J=5.4 Hz, 2 H), 4.13 (s, 3 H), 3.76 (t, J=5.1 Hz, 2 H), 3.34 (s, 3 H); MS (ESI): m/z: 339 [M+H]+.

Compound 9: (4-Methylthieno[3,2-b]pyrrol-5-yl)-[1-(2-phenoxyethyl)indol-3-yl]methanone

0.031 g of (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-(2-phenoxyethyl)indol-3-yl]methanone was prepared from 0.034 g (0.12 mmol) of 1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone (Compound 1) and 0.030 g (0.14 mmol) of 2-bromoethoxybenzene (Sigma-Aldrich, Cat. No. B75506-25G) according to the procedure described for Compound 7 (yield: 64%). 1H NMR (DMSO-d6) 5 (ppm): 8.31-8.17 (m, 2 H), 7.76-7.67 (m, 1 H), 7.60 (d, J=5.4 Hz, 1 H), 7.367.20 (m, 5 H), 7.12 (s, 1 H), 6.96-6.85 (m, 3 H), 4.71 (t, J=5.1 Hz, 2 H), 4.38 (t, J=5.1 Hz, 2 H), 4.02 (s, 3 H); MS (ESI): m/z: 401 [M+H]+.

Compound 10: (7-Hydroxy-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone

0.396 mL of a 1 M solution of TBAF in THF was added at RT to a solution of 0.158 g (0.380 mmol) of [7-[fert-butyl(dimethyl)silii]oxy-1 H -indol-3-yl]-(4-methylthieno[3,2-b]pyrrole-5-M)methanone in 3 mL of dry THF and stirred for 15 min. The solvent was then evaporated and the crude mixture was partitioned between EtOAc and water. The organic phase was dried over Na2SO4, filtered, concentrated, and purified by column chromatography (eluent: hexane/acetone, 0% to 30% acetone (by volume)) to afford 98 mg (yield: 86%) of (7-hydroxy-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone as a yellow solid. 1H r Mn (DMSO-d6) 5 (ppm): 11.91 (s a, 1 H), 9.86 (s a, 1 H), 7.92 (s, 1 H), 7.70-7.63 (m, 1 H), 7.56 (d, J=4.9 Hz, 1 H), 7.26 (d, J=4.9 Hz, 1 H), 7.12 (s, 1 H), 7.02-6.93 (m, 1 H), 6.68-6.58 (m, 1 H), 4.00 (s, 3 H); MS (ESI): m/z: 297 [M+H]+.

Compound 11: (4-Methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[2-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone hydrochloride

To a solution of 0.050 g (0.083 mmol) of tert-butyl 3-[[2-[2-[3-(4-methylthieno[3,2-b]pyrrole-5-carbonyl)indol-1-yl]ethoxy]phenoxy]methyl]pyrrolidine-1-carboxylate (Intermediate 23) dissolved in 1.0 mL of dioxane and 0.2 mL of MeOH was added 0.21 mL of 4 M HCl in dioxane. The mixture was stirred at RT for 6 h, then the solvent was removed, the residue was taken up first in MeOH and concentrated, and then in Et2O. After further concentration, the solid was triturated with Et2O/MeOH and dried to afford 0.042 g of (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[2-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]indol-3-yl]methanone hydrochloride (yield: 87%) as a brownish powder. 1H NMR (DMSO-d6) 5 (ppm): 8.75 (s a, 2 H), 8.30-8.16 (m, 2 H), 7.74-7.66 (m, 1 H), 7.60 (d, J=4.9 Hz, 1 H), 7.38-7.21 (m, 3 H), 7.13 (s, 1 H), 7.02-6.83 (m, 4 H), 4.71 (t, J=4.9 Hz, 2 H), 4.39 (t, J=4.9 Hz, 2 H), 4.02 (s, 3 H), 3.81-3.70 (m, 2 H), 3.21-3.03 (m, 2 H), 3.02-2.90 (m, 1 H), 2.88-2.76 (m, 1 H), 2.46-2.33 (m, 1 H), 1.87-1.75 (m, 1 H), 1.59-1.43 (m, 1 H); MS (ESI): m/z: 500 [M+H]+.

The following compounds were obtained from the corresponding Boc intermediates according to Compound 11 described in the procedure.

In the case of Compound 26, the compound was purified by preparative HPLC to afford the product as the trifluoroacetate salt. The solution was treated with saturated aqueous NaHCO3, and then the product was obtained by extraction with CH2Cl2, drying over Na2SO4 and removing the solvent in vacuo. Compounds 29 and 30 were obtained as free bases after filtration over a bicarbonate SPE cartridge (200 mg) and purification by column chromatography on silica gel (CH2Cl2/MeOH/NH3).

EXAMPLE 2. BIOLOGICAL TEST

2.1 KDM1A (LSD1) enzyme inhibition assay

KDM1A inhibitory activity was determined using a TR-FRET (time-resolved fluorescence resonance energy transfer) assay (Lance® Ultra Demethylase technology (Perkin Elmer, Waltham, MA, USA)), comprising a europium chelate donor dye (TRF0404, Perkin Elmer, Waltham, MA, USA) together withULight™ (TR0102, Perkin Elmer, Waltham, MA, USA), a small molecular weight acceptor dye with a red-shifted fluorescent emission, and a biotinylated 21-amino acid histone H3-derived monomethylated peptide (H3K4me) [Lys(Me1)4]-Histone<h>3 (1-21)-GGK(biotin), (64355, Anaspec, Fremont, CA, USA) as a substrate. The intensity of the light emission is proportional to the level of biotinylated reaction product. The recombinant human KDM1A/CoREST protein complex was produced in E. coli as separate proteins and copurified as previously described (Forneris, F. et al. Trends Biochem, Sci, 2008, 33, 181-189) (Forneris, F. et al. J. Biol. Chem. 2007, 282, 20070-20074).

Demethylase assay conditions: 0.25 nM KDM1A/CoREST protein and compound in 100% DMSO in a final volume of 48 µl of assay buffer (50 mM Tris HCl pH 8.8, 50 mM NaCl, 1 mM dTt, 0.01% Tween-20 (by volume)) were added to each well of a 96-well half-area white flat-bottom plate (3693 Costar, Sigma-Aldrich, St. Louis, M, USA).

The demethylase reaction was initiated by the addition of 50 nM monomethylated histone H3K4. After 20 min at RT, 300 gM tranylcypromine (P8511-1G, Sigma-Aldrich, St. Louis, MO 63103) was added to stop the reaction.

Detection step conditions: 10 µl of the assay mixture was transferred from the original plate to a white 384-well plate (6007290 OptiPlate™, Perkin Elmer, Waltham, MA, USA) and 10 µl of the detection mixture containing 2 nM Eu antibody and 10 nM U-Light-Streptavidin in 1X Lance Detection Buffer (TRF0404, TR0102, CR97100, Perkin Elmer, Waltham, MA, USA). The resulting mixture was incubated in the dark for 1 h at RT. The TR-FRET signal was then read by a fluorimeter (Infinite® F200, Tecan, Mannedorf, Switzerland) (excitation 320 nm, emission 665 nm and 620 nm, delay time 50 gs, window time 100 gs).

IC50 determination Inhibitor concentrations ranged from 0.025 to 500 gM (1:3 serial dilutions). The IC50 was calculated using GraphPad software.

Compounds 1-2,5-6 and 8 have IC50 values of less than 50 gM, Compound 11 has an IC50 value of less than 5 gM and Compounds 12-30 have IC50 values of less than 0.5 gM.

2.2 Cell growth

CellTiter-Flor® (Promega) is a non-lytic, single-reagent addition fluorescence assay that measures the relative number of live cells in a culture population following experimental manipulation. The CellTiter-Fluor™ Cell Viability Assay measures constitutive and conserved protease activity in living cells and therefore acts as a marker of cell viability.

Human leukemia cells, MV4-11 (obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen, ACC 102) or exponentially growing NB4 cells (obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen), were incubated for 48 h with different concentrations of the inhibitors. After 48 h a volume of CellTiter-Fluor® Reagent equal to one-fifth of the volume of the cell culture medium was added. The contents were mixed and incubated for at least 90 min at 37 °C to obtain a stable signal. Fluorescence was recorded using an excitation wavelength of 360 nm and emission at 535 nm. The IC50 was calculated using GraphPad software. Compounds 11 and 13-19 exhibit IC50 values of less than 10 gM against MV4-11 human leukemia cells, Compounds 11 and 13-24 exhibit IC50 values of less than 10 gM against NB4 human leukemia cells.

2.3 Bioluminescent coupled assay for monoamine oxidases (MAO-Glo assay)

The Promega MAO Glo assay (cat. V1402, Promega, Madison, WI) was used to measure the effect of inhibitors of MAO A and MAO B activity. Recombinant human MAO A and MAO B were expressed in Pichia pastoris and purified as published (Binda C. et al. Proc. Natl. Acad. Sci. USA, 2003, 9750-9755). The assay was performed at RT in 50 µl (25 µl reaction solution 25 µl detection reagent) in white half-area 96-well plates (cat. 3693, Corning, Corning, NY). Luminescence was measured after 20 min incubation in the dark using a microplate reader (Infinite F200, Tecan Group, Switzerland) with an integration time of 0.25 s per well. 50 nM MAO A or 125 nM MAO B were incubated with five different concentrations of inhibitor (0.004 gM to 100 gM) for 15 min at RT in Promega MAO buffer or Promega MAO buffer B (MAO Glo assay kit, catalog number V1402, Promega, Madison, WI). After 30 min of incubation, the reaction was stopped with Promega Detection Reagent. All compounds were tested twice and IC50 values were calculated using GraphPad Prism version 4.0 (GraphPad Software, San Diego, CA).

Compounds 13-14 and 17-18 were at least 10-fold more active against KDM1A (LSD1) compared to both MAO A and MAO B.

Claims (15)

CLAIMS 1. A compound of formula (I) Res hydrogen or L1-R5; R1, R2 are each hydrogen, OH, C1-C4 alkyl, C3-C7 cycloalkyl, aryl or heteroaryl; R3 is hydrogen or C1-C4 alkyl; L1 is a bond, -(CH2)j-Y- or -(CH2)k-, jes an integer from 2 to 6; kes an integer from 1 to 6; Yes oxygen; R<5> is C1-C4 alkyl or aryl, where the aryl is optionally substituted with one or two substituents chosen from halogen, C1-C6 alkyl or L2-R6; L2 is -(CH2)m- or -(CH2)n-W-(CH2)p-; R<6> is heterocyclyl, where the heterocyclyl is optionally substituted with C1-C6 alkyl; m, n, p are, independently, zero or an integer from 1 to 6; Wes oxygen, n H or CH2; or a stereoisomer or a pharmaceutically acceptable salt thereof. 2. The compound according to claim 1, wherein R3 is methyl or ethyl. 3. The compound according to claim 1 or 2, wherein R is hydrogen or L1-R5; R1, R2 are independently hydrogen, OH or C1-C4 alkyl; R3 is methyl or methoxy; L1 is a bond, -(CH2)2-Y- or -(CH2)k-; k is an integer from 1 to 4; Yes oxygen; R5 is C1-C4 alkyl or phenyl substituted with one or two substituents chosen from L2-R6; L2 is -W-(CH2)p-; R<6> is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl; w is zero or 1; W is oxygen; or a stereoisomer or a pharmaceutically acceptable salt thereof. 4. The compound according to claim 1 or 2, wherein Res is hydrogen or L1-R5; R1, R2 are independently H or methyl; R3 is methyl or methoxy; L1 is -(CH2)j-Y- or -(CH2)k-; jes 2; kes 3; Yes oxygen; R5 is methyl or phenyl substituted with one or two substituents chosen from L2-R6; L2 is -(CH2)n-W-(CH2)p-; nes 0; pes 0 or 1; Wes oxygen; or a stereoisomer or a pharmaceutically acceptable salt thereof; optionally wherein: ResL1-R5; R1, R2 are hydrogen; R5 is phenyl substituted with one or two substituents chosen from L2-R6; more optionally wherein R3 is methyl; L1 is -(CH2)j-Y-R5 is phenyl substituted with one substituentL2-R6; 5. The compound according to any one of claims 1-4, wherein R6 is piperidinyl or pyrrolidinyl. 6. A compound according to claim 1 selected from: 1H-indol-3-yl-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (5-methyl-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (5-ethyl-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (5-bromo-1H-indol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (4-ethylthieno[3,2-b]pyrrol-5-yl)-(1H-indol-3-yl)methanone; (1-methylindol-3-yl)-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; [1-(2-methoxyethyl)indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-(2-phenoxyethyl)indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[2-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]i ndol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]i ndol-3-yl]methanone; (4-methylthieno[3,2-b]pirrol-5-yl)-[1-[2-[3-[[(3S)-pirrolidin-3-yl]methoxy]phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrole-5-yl)-[1-[2-[3-[[(3R)-pyrrolidin-3-yl]methoxy]phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(pyrrolidin-3-ylmethoxy)phenoxy]ethyl]i ndol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(4-piperidyloxy)phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidyloxy)phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[3-(4-piperidylmethoxy)phenoxy]ethyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[2-[4-(4-piperidylmethoxy)phenoxy]ethyl]indol-3-yl]methanone; [1-[2-[3-(azepan-4-yloxy)phenoxy]ethyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; [1-[2-[4-(azepan-4-yloxy)phenoxy]ethyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(4-piperidylmethoxy)phenyl]propyl]indol-3-yl]methanone; (4-methylthieno[3,2-b]pirrol-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone; [1-[3-[4-(azepan-4-yloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; [1-[3-[3,4-bis(4-piperidylmethoxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; (4-ethylthieno[3,2-b]pyrrole-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone; (4-ethylthieno[3,2-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone; [5-ethyl-1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; and [5-methyl-1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]-(4-methylthieno[3,2-b]pyrrol-5-yl)methanone; or a stereoisomer or a pharmaceutically acceptable salt thereof. 7. A compound of formula (I) R1, R2 are hydrogen; R4 is methyl or methoxy; L1 is -(CH2)j-Y- or -(CH2)k-, jes 2; kes 3; Yes oxygen; R5 is phenyl substituted with one or two substituents chosen from L2-R6; L2 is -(CH2)n-W-(CH2)p-; R<6> is heterocyclyl, wherein the heterocyclyl is optionally substituted with C1-C6 alkyl; nes 0; pes 0 or 1; Wes oxygen; or a stereoisomer or a pharmaceutically acceptable salt thereof. 8. The compound of claim 7, wherein said compound is: (6-methylthieno[2,3-b]pyrrol-5-yl)-[1-[3-[4-(pyrrolidin-3-ylmethoxy)phenyl]propyl]indol-3-yl]methanone; either (6-methylthieno[2,3-b]pirrol-5-yl)-[1-[3-[4-(4-piperidyloxy)phenyl]propyl]indol-3-yl]methanone. 9. A pharmaceutical composition comprising a compound as defined in any one of claims 1-8 together with a pharmaceutically acceptable excipient and/or diluent. optionally wherein the pharmaceutical composition further comprises at least one therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors or a chemotherapeutic agent. 10. The pharmaceutical composition according to claim 9 in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable or infusible liquid, solutions, suspensions, emulsions, suppositories, ointments, creams, lotions, gels, pastes or transdermal administration devices. 11. The compound according to any one of claims 1-8 for use as a medicament. 12. The compound according to any one of claims 1-8 for use in: (a) a method of treatment and/or prevention of cancer, infectious diseases or a disease characterized by aberration of cellular energy metabolism, such as obesity; (b) a method of treatment and/or prevention of leukemia, non-small cell lung cancer, hepatocellular carcinoma or glioblastomas; or (c) a method of treatment and/or prevention of acute myeloid leukemia. 13. The compound for use according to claim 12, wherein the method further comprises administering a therapeutically effective amount of at least one therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, antiproliferative/antineoplastic agents, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, antiangiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors or a chemotherapeutic agent. 14. A process for obtaining a compound of formula (I) according to claim 1, wherein R is hydrogen, the process comprising the preparation of the acyl chloride of formula A2 by reacting the carboxylic acid of formula A1 with thionyl chloride, and the preparation of the indole anion A4 by reacting the indole A3 with methylmagnesium bromide, and the condensation of the acyl chloride of formula A2 with the indole anion A4 to obtain a compound of formula (I), as shown below: A is where A, R1, R2 and R3 are as defined in claim 1. 15. A process for obtaining a compound of formula (I) according to claim 1 or claim 7, wherein ResL1-R5, the process comprising the reaction of a compound of formulaB1 with a compound of formulaB2 in the presence of a base, as shown below: wherein ResL1-R5 and A, R1, R2, R3, R4, L1 and R5 are as defined in claim 1 or claim 7, and LG is a leaving group; optionally wherein LG is bromine.